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Cattier B, Guignard R, Martel I, Martel C, Simard-Bisson C, Larouche D, Guiraud B, Bessou-Touya S, Germain L. Bulge-Derived Epithelial Cells Isolated from Human Hair Follicles Using Enzymatic Digestion or Explants Result in Comparable Tissue-Engineered Skin. Int J Mol Sci 2025; 26:1852. [PMID: 40076477 PMCID: PMC11899990 DOI: 10.3390/ijms26051852] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/29/2024] [Revised: 02/12/2025] [Accepted: 02/17/2025] [Indexed: 03/14/2025] Open
Abstract
Hair follicle stem cells, located in the bulge region of the outer root sheath, are multipotent epithelial stem cells capable of differentiating into epidermal, sebaceous gland, and hair shaft cells. Efficient culturing of these cells is crucial for advancements in dermatology, regenerative medicine, and skin model development. This investigation aimed to develop a protocol for isolating enriched bulge-derived epithelial cells from scalp specimens to produce tissue-engineered substitutes. The epithelium, including hair follicles, was separated from the dermis using thermolysin, followed by microdissection of the bulge region. Epithelial stem cells were isolated using enzymatic dissociation to create a single-cell suspension and compared with the direct explant culture and a benchmark method which isolates cells from the epidermis and pilosebaceous units. After 8 days of culture, the enzymatic digestion of microdissected bulges yielded 5.3 times more epithelial cells compared to explant cultures and proliferated faster than the benchmark method. Cells cultured from all methods exhibited comparable morphology and growth rates. The fully stratified epidermis of tissue-engineered skin was similar, indicating comparable differentiation potential. This enzymatic digestion method improved early-stage cell recovery and expansion while maintaining keratinocyte functionality, offering an efficient hair bulge cell-extraction technique for tissue engineering and regenerative medicine applications.
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Affiliation(s)
- Bettina Cattier
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Rina Guignard
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Israël Martel
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Christian Martel
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Carolyne Simard-Bisson
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Danielle Larouche
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
| | - Béatrice Guiraud
- R&D Center, Pierre Fabre Dermo-Cosmétique, 31100 Toulouse, France; (B.G.); (S.B.-T.)
| | - Sandrine Bessou-Touya
- R&D Center, Pierre Fabre Dermo-Cosmétique, 31100 Toulouse, France; (B.G.); (S.B.-T.)
| | - Lucie Germain
- Centre de Recherche en Organogénèse Expérimentale de l’Université Laval/LOEX, Quebec City, QC G1J 5B3, Canada; (B.C.); (R.G.); (I.M.); (C.M.); (C.S.-B.); (D.L.)
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, QC G1V 0A6, Canada
- CHU de Québec-Université Laval Research Centre, Quebec City, QC G1J 5B3, Canada
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Zhang Y, Zhao X, Li S, Xu Y, Bai S, Zhang W. Melatonin-Mediated Circadian Rhythm Signaling Exhibits Bidirectional Regulatory Effects on the State of Hair Follicle Stem Cells. Biomolecules 2025; 15:226. [PMID: 40001528 PMCID: PMC11852975 DOI: 10.3390/biom15020226] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2024] [Revised: 01/27/2025] [Accepted: 01/29/2025] [Indexed: 02/27/2025] Open
Abstract
The development and regulation of hair are widely influenced by biological rhythm signals. Melatonin plays a crucial role as a messenger in transmitting biological rhythm signals, and its impact on hair development has been well documented. During the process of hair follicle reconstruction, hair follicle stem cells (HFSCs) are the most important cell type, but the regulatory effect of melatonin on the state of HFSCs is still not fully understood. Therefore, it is necessary to conduct a more comprehensive characterization of the effects of melatonin on the state of hair follicle stem cells. The research results indicate that HFSCs express retinoic acid receptor-related orphan receptor alpha (Rorα), and melatonin inhibits the expression level of RORA. Experimental results from CUT&Tag, CUT&RUN, and dual luciferase reporter assays demonstrate that Foxc1 is a downstream target gene of RORA, with RORA regulating Foxc1 expression by binding to the promoter region of Foxc1. The CCK-8 assay results show that low doses of melatonin upregulate the survival rate of hair follicle stem cells, while high doses have the opposite effect. The knockdown of Foxc1 reverses the inhibitory effect of high-dose melatonin on the survival rate of hair follicle stem cells. Based on these findings, we believe that melatonin-mediated circadian signals exert a bidirectional regulatory effect on the state of HFSCs.
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Affiliation(s)
- Yu Zhang
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
| | - Xuefei Zhao
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
- National Forestry and Grassland Administration Research Center of Engineering Technology for Wildlife Conservation and Utilization, Harbin 150040, China
| | - Shuqi Li
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
| | - Yanchun Xu
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
- National Forestry and Grassland Administration Research Center of Engineering Technology for Wildlife Conservation and Utilization, Harbin 150040, China
- Detecting Center of Wildlife, State Forestry and Grassland Administration, Harbin 150040, China
| | - Suying Bai
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
- Detecting Center of Wildlife, State Forestry and Grassland Administration, Harbin 150040, China
| | - Wei Zhang
- College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China; (Y.Z.); (X.Z.)
- Detecting Center of Wildlife, State Forestry and Grassland Administration, Harbin 150040, China
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Sousa P, Lopes B, Sousa AC, Coelho A, de Sousa Moreira A, Rêma A, Gonçalves-Maia M, Amorim I, Alvites R, Alves N, Geuna S, Maurício AC. Isolation, Expansion, and Characterization of Rat Hair Follicle Stem Cells and Their Secretome: Insights into Wound Healing Potential. Biomedicines 2024; 12:2854. [PMID: 39767760 PMCID: PMC11672956 DOI: 10.3390/biomedicines12122854] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/31/2024] [Revised: 11/23/2024] [Accepted: 12/11/2024] [Indexed: 01/11/2025] Open
Abstract
Background: Stem cells are capable of self-renewal and differentiation into various specialized cells, making them a potential therapeutic option in regenerative medicine. This study establishes a comprehensive methodology for isolating, culturing, and characterizing rat hair follicle stem cells. Methods and Results: Hair follicles were harvested from Sprague-Dawley rats and subjected to two different isolation techniques. Immunohistochemical analysis and real-time PCR confirm the expression of specific surface markers and genes, validating the cells' identity. Growth kinetics, colony formation units (CFU), and tri-differentiation capacity were also assessed. Additionally, the cells' secretome was analyzed, regarding its content in biofactors with wound healing properties. Conclusions: These findings highlight the potential of these cells as a valuable cell source for skin regeneration applications. They contribute to advancing our understanding of stem cell applications in regenerative medicine and hold promise for therapeutic interventions in various clinical contexts, aligning with broader research on the diverse capabilities of hair follicle stem cells.
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Affiliation(s)
- Patrícia Sousa
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - Bruna Lopes
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - Ana Catarina Sousa
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - André Coelho
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - Alícia de Sousa Moreira
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - Alexandra Rêma
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
| | - Maria Gonçalves-Maia
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Maia & Muller-Biotech, Rua Alfredo Allen, 455/461, 4200-135 Porto, Portugal
| | - Irina Amorim
- Departamento de Patologia e Imunologia Molecular, ICBAS—School of Medicine and Biomedical Sciences, University of Porto (UP), Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal;
- Institute for Research and Innovation in Health (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
- Institute of Molecular Pathology and Immunology, University of Porto (IPATIMUP), Rua Júlio Amaral de Carvalho 45, 4200-135 Porto, Portugal
| | - Rui Alvites
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
- Instituto Universitário de Ciências da Saúde (CESPU), Avenida Central de Gandra 1317, 4585-116 Paredes, Portugal
| | - Nuno Alves
- Centre for Rapid and Sustainable Product Development, Polytechnic of Leiria, 2430-028 Marinha Grande, Portugal;
| | - Stefano Geuna
- Department of Clinical and Biological Sciences, Cavalieri Ottolenghi Neuroscience Institute, University of Turin, Ospedale San Luigi, 10043 Turin, Italy;
| | - Ana Colette Maurício
- Departamento de Clínicas Veterinárias, Instituto de Ciências Biomédicas de Abel Salazar (ICBAS), Universidade do Porto (UP), Rua de Jorge Viterbo Ferreira, No. 228, 4050-313 Porto, Portugal; (P.S.); (B.L.); (A.C.S.); (A.C.); (A.d.S.M.); (A.R.); (R.A.)
- Centro de Estudos de Ciência Animal (CECA), Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto (ICETA), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal;
- Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal
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Mahjoor M, Nobakht M, Ataei Kachouei F, Zalpoor H, Heidari F, Yari A, Joulai Veijouye S, Nazari H, Sajedi N. In Vitro differentiation of hair-follicle bulge stem cells into synaptophysin-expressing neurons: a potential new approach for neuro-regeneration. Hum Cell 2024; 38:19. [PMID: 39546092 DOI: 10.1007/s13577-024-01146-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2024] [Accepted: 10/09/2024] [Indexed: 11/17/2024]
Abstract
Stem cells, particularly bulge hair follicle stem cells (HFSCs), have recently attracted significant interest due to their potential for tissue repair and regeneration. These cells, marked by their expression of Nestin (a neural stem cell marker), suggest the possibility of neural differentiation into neurons. This study investigated the use of retinoic acid (RA) and epidermal growth factor (EGF) to induce HFSC transformation into mature neurons, identified by synaptophysin expression. Rat whisker follicles were cultured in a medium suitable for HFSC survival and proliferation. Immunostaining techniques were used to identify HFSCs and assess their differentiation into neural cells. The addition of RA and EGF to the culture medium aimed to induce this differentiation. Findings demonstrate that HFSCs expressed Nestin, indicating their pluripotent nature. Treatment with RA and EGF resulted in synaptophysin expression, a marker of mature neurons, which was absent in the control group. However, this treatment group also displayed a decrease in the expression of other neural markers (βIII tubulin and NeuN). This study suggests that a combination of RA and EGF can accelerate HFSC differentiation into synaptophysin-positive cells in vitro. This research paves the way for further exploration of its potential application in neuro-regeneration.
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Affiliation(s)
- Mohamad Mahjoor
- Cellular and Molecular Research Centre, Qom University of Medical Sciences, Qom, Iran
- Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Maliheh Nobakht
- Department of Anatomy, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | | | - Hamidreza Zalpoor
- Shiraz Neuroscience Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
- Network of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Education & Research Network (USERN), Universal Scientific, Tehran, Iran.
| | - Fatemeh Heidari
- Department of Anatomy, Faculty of Medicine, Qum University of Medical Sciences, Qom, Iran
| | - Abazar Yari
- Department of Anatomy, School of Medicine, Alborz University of Medical Science, Karaj, Iran
| | | | - Hojjatollah Nazari
- School of Biomedical Engineering, Faculty of Engineering and IT, University of Technology Sydney, Sydney, NSW, Australia
| | - Nayereh Sajedi
- Department of Anatomy, Faculty of Medicine, Qom Medical Sciences, Islamic Azad University, Qom, Iran.
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Pandamooz S, Chavoshinezhad S, Mostaghel M, Rasekh A, Ghorbani N, Dara M, Pandamooz T, Tanideh N, Salehi MS. Directing Rat Hair Follicle Stem Cells Toward Neuronal Lineage With Enhanced Trophic Factor Expression. Adv Biomed Res 2024; 13:84. [PMID: 39512401 PMCID: PMC11542700 DOI: 10.4103/abr.abr_111_24] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2024] [Revised: 04/30/2024] [Accepted: 05/05/2024] [Indexed: 11/15/2024] Open
Abstract
Background Hair follicle stem cells (HFSCs) are promising candidates for cell-based therapies in neurodegenerative diseases because of their ability to differentiate into neural lineages and exert paracrine effects in damaged tissues. However, their clinical application faces challenges, particularly in efficiently guiding them toward neural lineages. This study explores using chick embryo extract (CEE) to enhance HFSCs' secretory capacity and neuronal differentiation. Materials and Methods HFSCs from rat whisker pads were cultured in growth medium supplemented with either 20% FBS or a combination of 10% FBS and 10% CEE, transitioning to 20% FBS after the first subculture. We conducted gene expression profiling of lineage commitment markers and neurotrophic factors in both experimental groups, alongside morphological assessments and protein expression analyses. Results CEE supplementation during migration increased neuronal differentiation, evidenced by more cells with neurites and higher MAP2 expression at both the gene and protein levels. CEE also inhibited the expression of PDGFR-α, indicating a suppression of differentiation toward Schwann cells. Furthermore, we observed increased levels of trophic factors such as BDNF and VEGF at passage 3 induced by CEE supplementation. Conclusions Enhancing the neuronal lineage commitment of hair follicle stem cells (HFSCs) and boosting the expression of trophic and angiogenic factors through short-term CEE preconditioning during their migratory stage presents a compelling approach. This strategy holds great promise in enhancing the effectiveness of stem cell-based therapies for neurological disorders.
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Affiliation(s)
- Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Sara Chavoshinezhad
- Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran
| | - Mandana Mostaghel
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Armita Rasekh
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Nasrin Ghorbani
- Department of Nursing, College of Nursing, Lebanese French University, Erbil, Kurdistan, Iraq
| | - Mahintaj Dara
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Tahoura Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Nader Tanideh
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mohammad Saied Salehi
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
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Dermitzakis I, Kampitsi DD, Manthou ME, Evangelidis P, Vakirlis E, Meditskou S, Theotokis P. Ontogeny of Skin Stem Cells and Molecular Underpinnings. Curr Issues Mol Biol 2024; 46:8118-8147. [PMID: 39194698 DOI: 10.3390/cimb46080481] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2024] [Revised: 07/21/2024] [Accepted: 07/25/2024] [Indexed: 08/29/2024] Open
Abstract
Skin stem cells (SCs) play a pivotal role in supporting tissue homeostasis. Several types of SCs are responsible for maintaining and regenerating skin tissue. These include bulge SCs and others residing in the interfollicular epidermis, infundibulum, isthmus, sebaceous glands, and sweat glands. The emergence of skin SCs commences during embryogenesis, where multipotent SCs arise from various precursor populations. These early events set the foundation for the diverse pool of SCs that will reside in the adult skin, ready to respond to tissue repair and regeneration demands. A network of molecular cues regulates skin SC behavior, balancing quiescence, self-renewal, and differentiation. The disruption of this delicate equilibrium can lead to SC exhaustion, impaired wound healing, and pathological conditions such as skin cancer. The present review explores the intricate mechanisms governing the development, activation, and differentiation of skin SCs, shedding light on the molecular signaling pathways that drive their fate decisions and skin homeostasis. Unraveling the complexities of these molecular drivers not only enhances our fundamental knowledge of skin biology but also holds promise for developing novel strategies to modulate skin SC fate for regenerative medicine applications, ultimately benefiting patients with skin disorders and injuries.
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Affiliation(s)
- Iasonas Dermitzakis
- Department of Histology-Embryology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
| | - Despoina Dimitria Kampitsi
- Department of Histology-Embryology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
| | - Maria Eleni Manthou
- Department of Histology-Embryology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
| | - Paschalis Evangelidis
- Hematology Unit-Hemophilia Centre, 2nd Propedeutic Department of Internal Medicine, Hippocration Hospital, Aristotle University of Thessaloniki, 54642 Thessaloniki, Greece
| | - Efstratios Vakirlis
- First Department of Dermatology and Venereology, School of Medicine, Aristotle University of Thessaloniki, 54643 Thessaloniki, Greece
| | - Soultana Meditskou
- Department of Histology-Embryology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
| | - Paschalis Theotokis
- Department of Histology-Embryology, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
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Hasegawa-Haruki A, Obara K, Takaoka N, Shirai K, Hamada Y, Arakawa N, Aki R, Hoffman RM, Amoh Y. Hair-follicle associated pluripotent (HAP)-cell-sheet implantation enhanced wound healing in diabetic db/db mice. PLoS One 2024; 19:e0304676. [PMID: 38875234 PMCID: PMC11178214 DOI: 10.1371/journal.pone.0304676] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2023] [Accepted: 05/15/2024] [Indexed: 06/16/2024] Open
Abstract
Diabetes often results in chronic ulcers that fail to heal. Effective treatment for diabetic wounds has not been achieved, although stem-cell-treatment has shown promise. Hair-follicle-associated-pluripotent (HAP)-stem-cells from bulge area of mouse hair follicle have been shown to differentiate into keratinocytes, vascular endothelial cells, smooth muscle cells, and some other types of cells. In the present study, we developed HAP-cell-sheets to determine their effects on wound healing in type-2 diabetes mellitus (db/db) C57BL/6 mouse model. Flow cytometry analysis showed cytokeratin 15 expression in 64% of cells and macrophage expression in 3.6% of cells in HAP-cell-sheets. A scratch cell migration assay in vitro showed the ability of fibroblasts to migrate and proliferate was enhanced when co-cultured with HAP-cell-sheets. To investigate in vivo effects of the HAP-cell-sheets, they were implanted into 10 mm circular full-thickness resection wounds made on the back of db/db mice. Wound closure was facilitated in the implanted group until day 16. The thickness of epithelium and granulation tissue volume at day 7 were significantly increased by the implantation. CD68 positive area and TGF-β1 positive area were significantly increased; meanwhile, iNOS positive area was reduced at day 7 in the HAP-cell-sheets implanted group. After 21 days, CD68 positive areas in the implanted group were reduced to under the control group level, and TGF-β1 positive area had no difference between the two groups. These observations strongly suggest that the HAP-cell-sheets implantation is efficient to facilitate early macrophage activity and to suppress inflammation level. Using immuno-double-staining against CD34 and α-SMA, we found more vigorous angiogenesis in the implanted wound tissue. The present results suggest autologous HAP-cell-sheets can be used to heal refractory diabetic ulcers and have clinical promise.
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Affiliation(s)
- Ayami Hasegawa-Haruki
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Sagamihara, Kanagawa, Japan
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Koya Obara
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nanako Takaoka
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Sagamihara, Kanagawa, Japan
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Kyoumi Shirai
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nobuko Arakawa
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Ryoichi Aki
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Robert M Hoffman
- AntiCancer, Inc., San Diego, California, United states of America
- Department of Surgery, University of California San Diego, San Diego, California, United states of America
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Sagamihara, Kanagawa, Japan
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
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8
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Kubota Y, Varshney N, Kobayashi K, Tsunoda T, Hoffman RM. Methionine Dependence of Hair Maintenance in C57BL/6 Mice. In Vivo 2024; 38:1199-1202. [PMID: 38688645 PMCID: PMC11059892 DOI: 10.21873/invivo.13555] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2023] [Revised: 01/30/2024] [Accepted: 01/31/2024] [Indexed: 05/02/2024]
Abstract
BACKGROUND/AIM Hair-follicle keratinocytes contain high levels of cysteine, which is derived from methionine, rapidly proliferate, and form the hair shaft. The high proliferation rate of hair-follicle keratinocytes resembles that of aggressive cancer cells. In the present study, we determined the effect of a methionine-deficient diet on hair loss (alopecia) in mice with or without homocysteine supplementation. MATERIALS AND METHODS Mice were fed a normal rodent diet (2020X, ENVIGO) (Group 1); a methionine-choline-deficient diet (TD.90262, ENVIGO) (Group 2); a methionine-choline-deficient diet with a 10 mg/kg/day supply of homocysteine administered by intra-peritoneal (i.p.) injection for 2 weeks (Group 3). In Group 2, mice were fed a methionine-choline-deficient diet for an additional 2 weeks but with 10 mg/kg/day of i.p. l-homocysteine and the mice were observed for two additional weeks. Subsequently, the mice were fed a standard diet that included methionine. Hair loss was monitored by photography. RESULTS After 14 days, hair loss was observed in Group 2 mice on a methionine-restricted diet but not in Group 3 mice on the methionine-restricted diet which received i.p. homocysteine. In Group 2, at 2 weeks after methionine restriction, hair loss was not rescued by homocysteine supplementation. However, after restoration of methionine in the diet, hair growth resumed. Thus, after 2 weeks of methionine restriction, only methionine restored hair loss, not homocysteine. CONCLUSION Hair maintenance requires methionine in the diet. Future experiments will determine the effects of methionine restriction on hair-follicle stem cells.
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Affiliation(s)
- Yutaro Kubota
- AntiCancer Inc., San Diego, CA, U.S.A
- Department of Surgery, University of California, San Diego, CA, U.S.A
- Division of Internal Medicine, Department of Medical Oncology, Showa University School of Medicine, Tokyo, Japan
| | | | | | - Takuya Tsunoda
- Division of Internal Medicine, Department of Medical Oncology, Showa University School of Medicine, Tokyo, Japan
| | - Robert M Hoffman
- AntiCancer Inc., San Diego, CA, U.S.A.;
- Department of Surgery, University of California, San Diego, CA, U.S.A
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9
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Takaoka N, Yamane M, Hasegawa A, Obara K, Shirai K, Aki R, Hatakeyama H, Hamada Y, Arakawa N, Tanaka M, Hoffman RM, Amoh Y. Rat hair-follicle-associated pluripotent (HAP) stem cells can differentiate into atrial or ventricular cardiomyocytes in culture controlled by specific supplementation. PLoS One 2024; 19:e0297443. [PMID: 38277391 PMCID: PMC10817212 DOI: 10.1371/journal.pone.0297443] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/05/2023] [Accepted: 01/04/2024] [Indexed: 01/28/2024] Open
Abstract
There has been only limited success to differentiate adult stem cells into cardiomyocyte subtypes. In the present study, we have successfully induced beating atrial and ventricular cardiomyocytes from rat hair-follicle-associated pluripotent (HAP) stem cells, which are adult stem cells located in the bulge area. HAP stem cells differentiated into atrial cardiomyocytes in culture with the combination of isoproterenol, activin A, bone morphogenetic protein 4 (BMP4), basic fibroblast growth factor (bFGF), and cyclosporine A (CSA). HAP stem cells differentiated into ventricular cardiomyocytes in culture with the combination of activin A, BMP4, bFGF, inhibitor of Wnt production-4 (IWP4), and vascular endothelial growth factor (VEGF). Differentiated atrial cardiomyocytes were specifically stained for anti-myosin light chain 2a (MLC2a) antibody. Ventricular cardiomyocytes were specially stained for anti-myosin light chain 2v (MLC2v) antibody. Quantitative Polymerase Chain Reaction (qPCR) showed significant expression of MLC2a in atrial cardiomyocytes and MLC2v in ventricular cardiomyocytes. Both differentiated atrial and ventricular cardiomyocytes showed characteristic waveforms in Ca2+ imaging. Differentiated atrial and ventricular cardiomyocytes formed long myocardial fibers and beat as a functional syncytium, having a structure similar to adult cardiomyocytes. The present results demonstrated that it is possible to induce cardiomyocyte subtypes, atrial and ventricular cardiomyocytes, from HAP stem cells.
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Affiliation(s)
- Nanako Takaoka
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Michiko Yamane
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Ayami Hasegawa
- Department of Dermatology, Kitasato University Graduate School of Medical Sciences, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Koya Obara
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Kyoumi Shirai
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Ryoichi Aki
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Hiroyasu Hatakeyama
- Department of Physiology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Nobuko Arakawa
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Manabu Tanaka
- Bio-Imaging Center, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
| | - Robert M. Hoffman
- AntiCancer, Inc., San Diego, CA, United States of America
- Department of Surgery, University of California, San Diego, CA, United States of America
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University School of Medicine, Kitasato University School of Medicine, Minami Ward, Sagamihara, Japan
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10
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Akbari S, Haghani M, Ghobadi M, Hooshmandi E, Haghighi AB, Salehi MS, Pandamooz S, Azarpira N, Afshari A, Zabihi S, Nemati M, Bayat M. Combination Therapy with Platelet-Rich Plasma and Epidermal Neural Crest Stem Cells Increases Treatment Efficacy in Vascular Dementia. Stem Cells Int 2023; 2023:3784843. [PMID: 38146481 PMCID: PMC10749736 DOI: 10.1155/2023/3784843] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2023] [Revised: 11/07/2023] [Accepted: 11/28/2023] [Indexed: 12/27/2023] Open
Abstract
This study aimed to evaluate the efficacy and treatment mechanism of platelet-rich plasma (PRP) and neural crest-derived epidermal stem cells (ESCs) in their administration alone and combination in vascular dementia (VaD) model by two-vessel occlusion (2VO). Methods. Sixty-six rats were divided into six groups: the control, sham, 2VO + vehicle, 2VO + PRP, 2VO + ESC, and 2VO + ESC + PRP. The treated groups received 1 million cells on days 4, 14, and 21 with or without 500 µl PRP (twice a week) after 2VO. The memory performance and anxiety were evaluated by behavioral tests including open field, passive avoidance, and Morris water maze. The basal-synaptic transmission (BST) and long-term potentiation (LTP) were assessed through field-potential recordings of the CA1. The mRNA expression levels of IGF-1, TGF-β1, PSD-95, and GSk-3β were measured in the rat hippocampus by quantitative reverse transcription polymerase chain reaction. Results. The results demonstrated impaired learning, memory, and synaptic plasticity in the 2VO rats, along with a significant decrease in the expression of IGF-1, TGF-β1, PSD-95, and upregulation of GSK-3β. Treatment with ESC alone and ESC + PRP showed similar improvements in spatial memory and LTP induction, with associated upregulation of PSD-95 and downregulation of GSK-3β. However, only the ESC + PRP group showed recovery in BST. Furthermore, combination therapy was more effective than PRP monotherapy for LTP and memory. Conclusions. The transplantation of ESC showed better effects than PRP alone, and combination therapy increased the treatment efficacy with the recovery of BST. This finding may be a clue for the combination therapy of ESC and PRP for VaD.
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Affiliation(s)
- Somayeh Akbari
- Histomorphometry and Stereology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Masoud Haghani
- Histomorphometry and Stereology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mojtaba Ghobadi
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Etrat Hooshmandi
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
| | | | - Mohammad Saied Salehi
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Negar Azarpira
- Shiraz Institute of Stem Cell and Regenerative Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Afsoon Afshari
- Shiraz Nephro-Urology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Shahrbanoo Zabihi
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Marzieh Nemati
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mahnaz Bayat
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
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11
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Hooshmandi E, Akbari S, Pandamooz S, Ghobadi M, Ghasemi R, Maghsoudi N, Rai SN, Borhani-Haghighi A, Salehi MS, Azarpira N, YousefiNejad A, Haghani M, Bayat M. Combined use of hair follicle stem cells and CEPO (carbamylated erythropoietin)-Fc in a rat model of chronic cerebral hypoperfusion: A behavioral, electrophysiological, and molecular study. Behav Brain Res 2023; 454:114655. [PMID: 37666305 DOI: 10.1016/j.bbr.2023.114655] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2023] [Revised: 08/22/2023] [Accepted: 08/31/2023] [Indexed: 09/06/2023]
Abstract
BACKGROUND In dementia, synaptic dysfunction appears before neuronal loss. Stem cell therapy could potentially provide a promising strategy for the treatment of dementia models. The carbamylated erythropoietin fusion protein (CEPO-Fc) has shown synaptotrophic effects. This study aimed to determine the efficiency of the combined use of hair follicle stem cells (HFSC) and CEPO-Fc in the basal synaptic transmission (BST) and long-term plasticity (LTP) of chronic cerebral hypoperfusion (CCH) rats. METHODS We divided 64 adult rats into control, sham, CCH+vehicle, CCH+CEPO, CCH+HFSC, and CCH+HFSC+CEPO groups. The CEPO-Fc was injected three times/week for 30 days. HFSC transplantation was done on days 4, 14, and 21 after surgery. The Morris water maze test and passive avoidance were used to assess memory. BST and LTP were assessed by a field-potential recording of the CA1 region. The hippocampal mRNA expression of IGF-1, TGF-β1, β1-Catenine, NR2B, PSD-95, and GSk-3β was evaluated by quantitative RT-PCR. RESULTS Following combination therapy, spatial memory retention, and BST showed significant improvement relative to HFSC and CEPO-Fc groups. These effects were also confirmed by recovered mRNA expression of β1-catenin, TGF-β1, and NR2B. GSK-3β expression was downregulated in all treatment groups. The upregulated PSD-95 was identified in HFSC and combination groups compared to the vehicle group. CONCLUSIONS These findings indicate that the combined use of HFSC and CEPO-Fc may be more advantageous for treating memory disruption in the CCH model than CEPO-Fc or HFSC alone. This type of combination therapy may hopefully lead to a new approach to treatment for dementia.
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Affiliation(s)
- Etrat Hooshmandi
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Somayeh Akbari
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran; Histomorphometry and Stereology Research Centre, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Mojtaba Ghobadi
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Rasoul Ghasemi
- Neurophysiology Research Center and Physiology Department, Shahid Beheshti University of Medical Sciences, Tehran, the Islamic Republic of Iran
| | - Nader Maghsoudi
- Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, the Islamic Republic of Iran
| | | | - Afshin Borhani-Haghighi
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Mohammad Saied Salehi
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Negar Azarpira
- Transplant Research Center, Shiraz University of Medical Sciences, Mohammad Rasoul-Allah Research Tower, Shiraz, the Islamic Republic of Iran
| | - Amirhossein YousefiNejad
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Masoud Haghani
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran; Histomorphometry and Stereology Research Centre, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran
| | - Mahnaz Bayat
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, the Islamic Republic of Iran.
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12
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Pandamooz S, Salehi MS, Jurek B, Meinung CP, Azarpira N, Dianatpour M, Neumann ID. Oxytocin Receptor Expression in Hair Follicle Stem Cells: A Promising Model for Biological and Therapeutic Discovery in Neuropsychiatric Disorders. Stem Cell Rev Rep 2023; 19:2510-2524. [PMID: 37548806 DOI: 10.1007/s12015-023-10603-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 07/27/2023] [Indexed: 08/08/2023]
Abstract
The intricate nature of the human brain and the limitations of existing model systems to study molecular and cellular causes of neuropsychiatric disorders represent a major challenge for basic research. The promising progress in patient-derived stem cell technology and in our knowledge on the role of the brain oxytocin (OXT) system in health and disease offer new possibilities in that direction. In this study, the rat hair follicle stem cells (HFSCs) were isolated and expanded in vitro. The expression of oxytocin receptors (OXTR) was evaluated in these cells. The cellular viability was assessed 12 h post stimulation with OXT. The activation of OXTR-coupled intracellular signaling cascades, following OXT treatment was determined. Also, the influence of OXT on neurite outgrowth and cytoskeletal rearrangement were defined. The assessment of OXTR protein expression revealed this receptor is expressed abundantly in HFSCs. As evidenced by the cell viability assay, no adverse or cytotoxic effects were detected following 12 h treatment with different concentrations of OXT. Moreover, OXTR stimulation by OXT resulted in ERK1/2, CREB, and eEF2 activation, neurite length alterations, and cytoskeletal rearrangements that reveal the functionality of this receptor in HFSCs. Here, we introduced the rat HFSCs as an easy-to-obtain stem cell model that express functional OXTR. This cell-based model can contribute to our understanding of the progression and treatment of neuropsychiatric disorders with oxytocinergic system deficiency.
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Affiliation(s)
- Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Department of Molecular and Behavioural Neurobiology, University of Regensburg, Regensburg, Germany
| | - Mohammad Saied Salehi
- Department of Molecular and Behavioural Neurobiology, University of Regensburg, Regensburg, Germany.
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
| | - Benjamin Jurek
- Department of Molecular and Behavioural Neurobiology, University of Regensburg, Regensburg, Germany
- Research Group Neurobiology of Stress Resilience, Max Planck Institute of Psychiatry, Munich, Germany
| | - Carl-Philipp Meinung
- Department of Molecular and Behavioural Neurobiology, University of Regensburg, Regensburg, Germany
| | - Negar Azarpira
- Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mehdi Dianatpour
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Inga D Neumann
- Department of Molecular and Behavioural Neurobiology, University of Regensburg, Regensburg, Germany.
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13
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Tang H, Zhang X, Hao X, Dou H, Zou C, Zhou Y, Li B, Yue H, Wang D, Wang Y, Yang C, Fu J. Hepatocyte growth factor-modified hair follicle stem cells ameliorate cerebral ischemia/reperfusion injury in rats. Stem Cell Res Ther 2023; 14:25. [PMID: 36782269 PMCID: PMC9926795 DOI: 10.1186/s13287-023-03251-5] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2021] [Accepted: 08/22/2022] [Indexed: 02/15/2023] Open
Abstract
BACKGROUND Hair follicle stem cells (HFSCs) are considered as a promising cell type in the stem cell transplantation treatment of neurological diseases because of their rich sources, easy access, and the same ectoderm source as the nervous system. Hepatocyte growth factor (HGF) is a pleiotropic cytokine that shows neuroprotective function in ischemic stroke. Here we assessed the therapeutic effects of HFSCs on ischemic stroke injury and the synthetic effect of HGF along with HFSCs. METHODS Rat HFSCs were intravenously transplanted into a middle cerebral artery ischemia/reperfusion (I/R) rat model. Neurological scoring and TTC staining were performed to assess the benefits of HFSC transplantation. Inflammatory cytokines, blood-brain barrier integrity and angiogenesis within penumbra were estimated by Western blot and immunohistochemistry. The differentiation of HFSCs was detected by immunofluorescence method 2 weeks after transplantation. RESULTS HFSC transplantation could significantly inhibit the activation of microglia, improve the integrity of blood-brain barrier and reduce brain edema. Moreover, the number of surviving neurons and microvessels density in the penumbra were upregulated by HFSC transplantation, leading to better neurological score. The combination of HFSCs and HGF could significantly improve the therapeutic benefit. CONCLUSION Our results indicate for the first time that HGF modified HFSCs can reduce I/R injury and promote the neurological recovery by inhibiting inflammatory response, protecting blood-brain barrier and promoting angiogenesis.
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Affiliation(s)
- Hao Tang
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Xuemei Zhang
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Xiaojun Hao
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Haitong Dou
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Chendan Zou
- grid.410736.70000 0001 2204 9268Department of Biochemistry and Molecular Biology, Harbin Medical University, No.157 Baojian Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Yinglian Zhou
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Bing Li
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Hui Yue
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Duo Wang
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Yifei Wang
- grid.412463.60000 0004 1762 6325Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086 Heilongjiang China
| | - Chunxiao Yang
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang, China.
| | - Jin Fu
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang, China.
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14
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Human-derived hair follicle stem cells and hydrogen sulfide on focal cerebral ischemia model: A comparative evaluation of radiologic, neurobehavioral and immunohistochemical results. Brain Res 2023; 1799:148170. [PMID: 36410427 DOI: 10.1016/j.brainres.2022.148170] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/20/2022] [Revised: 11/10/2022] [Accepted: 11/15/2022] [Indexed: 11/19/2022]
Abstract
The present study investigated the effects of intracerebral human-derived hair follicle stem cells (HFBSCs), whether alone or in combination with hydrogen sulfide (H2S) in a rat model of focal cerebral ischemia. The rats were randomly assigned into 4 groups (n = 10): Control (phosphate buffered saline (PBS)), Group A (at 24 h post-middle cerebral artery occlusion(MCAo), stereotaxic intracerebral, 1,0 × 106, total 10 μL HFBSCs), Group B (3-14 d post-MCAo, intraperitoneal (i.p.), 25 μM/kg/day H2S), Group AB (HFBSCs + H2S). Cranial magnetic resonance images were recorded on postoperative 1st and 28th days. Three dimensional analysis was performed to calculate the infarct volumes. Rotarod and cylinder tests were performed after MCAo and finally all rats were euthanized by cardiac perfusion at 28 days after MCAo for immunohistochemical analysis. The reduction in infarct volumes of rats receiving HFBSC was significant. The cranial infarct volume on the postoperative 28th day was significantly higher in the group in which H2S was administered alone compared to the HFBSC alone group. All animals showed steadily improved spontaneous locomotor activity from day 7 post-MCAo on rotarod test, from day 1 on cylinder test, but showed no significant differences at all times. In all groups, the grading scores of CD34, CD5, CD11b and GFAP immunohistochemical markers did not differ significantly. In conclusion, intracerebral HFBSC treatment after 24 h of ischemic stroke may be an effective way to reduce the cranial infarct volume, whereas H2S treatment alone or in combination with HFBSC may not be sufficient for ischemic brain injury.
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15
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Pandamooz S, Jurek B, Dianatpour M, Haerteis S, Limm K, Oefner PJ, Dargahi L, Borhani-Haghighi A, Miyan JA, Salehi MS. The beneficial effects of chick embryo extract preconditioning on hair follicle stem cells: A promising strategy to generate Schwann cells. Cell Prolif 2023:e13397. [PMID: 36631409 DOI: 10.1111/cpr.13397] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/24/2022] [Revised: 12/06/2022] [Accepted: 12/28/2022] [Indexed: 01/13/2023] Open
Abstract
The beneficial effects of hair follicle stem cells in different animal models of nervous system conditions have been extensively studied. While chick embryo extract (CEE) has been used as a growth medium supplement for these stem cells, this is the first study to show the effect of CEE on them. The rat hair follicle stem cells were isolated and supplemented with 10% fetal bovine serum plus 10% CEE. The migration rate, proliferative capacity and multipotency were evaluated along with morphometric alteration and differentiation direction. The proteome analysis of CEE content identified effective factors of CEE that probably regulate fate and function of stem cells. The CEE enhances the migration rate of stem cells from explanted bulges as well as their proliferation, likely due to activation of AP-1 and translationally controlled tumour protein (TCTP) by thioredoxin found in CEE. The increased length of outgrowth may be the result of cyclic AMP response element binding protein (CREB) phosphorylation triggered by active CamKII contained in CEE. Further, CEE supplementation upregulates the expression of vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor. The elevated expression of target genes and proteins may be due to CREB, AP-1 and c-Myc activation in these stem cells. Given the increased transcript levels of neurotrophins, VEGF, and the expression of PDGFR-α, S100B, MBP and SOX-10 protein, it is possible that CEE promotes the fate of these stem cells towards Schwann cells.
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Affiliation(s)
- Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Benjamin Jurek
- Research Group Neurobiology of Stress Resilience, Max Planck Institute of Psychiatry, Munich, Germany.,Institute of Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany
| | - Mehdi Dianatpour
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Silke Haerteis
- Institute of Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany
| | - Katharina Limm
- Institute of Functional Genomics, University of Regensburg, Regensburg, Germany
| | - Peter J Oefner
- Institute of Functional Genomics, University of Regensburg, Regensburg, Germany
| | - Leila Dargahi
- Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | | | - Jaleel A Miyan
- Faculty of Biology, Medicine & Health, Division of Neuroscience & Experimental Psychology, The University of Manchester, Manchester, UK
| | - Mohammad Saied Salehi
- Clinical Neurology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
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16
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Obara K, Shirai K, Hamada Y, Arakawa N, Hasegawa A, Takaoka N, Aki R, Hoffman RM, Amoh Y. Direct implantation of hair-follicle-associated pluripotent (HAP) stem cells repairs intracerebral hemorrhage and reduces neuroinflammation in mouse model. PLoS One 2023; 18:e0280304. [PMID: 36638123 PMCID: PMC9838830 DOI: 10.1371/journal.pone.0280304] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/14/2022] [Accepted: 12/27/2022] [Indexed: 01/14/2023] Open
Abstract
Intracerebral hemorrhage (ICH) is a leading cause of mortality with ineffective treatment. Hair-follicle-associated pluripotent (HAP) stem cells can differentiate into neurons, glial cells and many other types of cells. HAP stem cells have been shown to repair peripheral-nerve and spinal-cord injury in mouse models. In the present study, HAP stem cells from C57BL/6J mice were implanted into the injured brain of C57BL/6J or nude mice with induced ICH. After allo transplantation, HAP stem cells differentiated to neurons, astrocytes, oligodendrocytes, and microglia in the ICH site of nude mice. After autologous transplantation in C57BL/6J mice, HAP stem cells suppressed astrocyte and microglia infiltration in the injured brain. The mRNA expression levels of IL-10 and TGF-β1, measured by quantitative Real-Time RT-PCR, in the brain of C57BL/6J mice with ICH was increased by HAP-stem-cell implantation compared to the non-implanted mice. Quantitative sensorimotor function analysis, with modified limb-placing test and the cylinder test, demonstrated a significant functional improvement in the HAP-stem-cell-implanted C57BL/6J mice, compared to non-implanted mice. HAP stem cells have critical advantages over induced pluripotent stem cells, embryonic stem cells as they do not develop tumors, are autologous, and do not require genetic manipulation. The present study demonstrates future clinical potential of HAP-stem-cell repair of ICH, currently a recalcitrant disease.
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Affiliation(s)
- Koya Obara
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Kyoumi Shirai
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nobuko Arakawa
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Ayami Hasegawa
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nanako Takaoka
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Ryoichi Aki
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Robert M. Hoffman
- AntiCancer, Inc., San Diego, California, United States of America
- Department of Surgery, University of California San Diego, San Diego, California, United States of America
- * E-mail: (YA); (RMH)
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
- * E-mail: (YA); (RMH)
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17
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Han J, Lin K, Choo H, Chen Y, Zhang X, Xu RH, Wang X, Wu Y. Distinct bulge stem cell populations maintain the pilosebaceous unit in a β-catenin-dependent manner. iScience 2022; 26:105805. [PMID: 36619975 PMCID: PMC9813789 DOI: 10.1016/j.isci.2022.105805] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2022] [Revised: 11/21/2022] [Accepted: 12/09/2022] [Indexed: 12/14/2022] Open
Abstract
The pilosebaceous unit (PSU) is composed of multiple compartments and the self-renewal of PSU depends on distinct hair follicle stem cell (HFSC) populations. However, the differential roles of the HFSCs in sebaceous gland (SG) renewal have not been completely understood. Here, we performed multiple lineage tracing analysis to unveil the contribution of different HFSC populations to PSU regeneration during the hair cycle and wound healing. Our results indicated that the upper bulge stem cells contributed extensively to the SG replenishment during hair cycling, while HFSCs in the lower bugle did not. During skin wound healing, all HFSC populations participated in the SG replenishment. Moreover, β-catenin activation promoted the contribution of HFSCs to SG replenishment, whereas β-catenin deletion substantially repressed the event. Thus, our findings indicated that HFSCs contributed to SG replenishment in a β-catenin-dependent manner.
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Affiliation(s)
- Jimin Han
- School of Life Sciences, Tsinghua University, Beijing, China
| | - Kaijun Lin
- School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou, China
| | - HuiQin Choo
- Tsinghua-Berkeley Shenzhen Institute (TBSI), Tsinghua University, Shenzhen, China
| | - Yu Chen
- School of Life Sciences, Tsinghua University, Beijing, China
- State Key Laboratory of Chemical Oncogenomics, and Shenzhen Key Laboratory of Health Sciences and Technology, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, China
| | - Xuezheng Zhang
- School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou, China
| | - Ren-He Xu
- Faculty of Health Sciences, University of Macau, Macau, China
| | - Xusheng Wang
- School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou, China
- Corresponding author
| | - Yaojiong Wu
- School of Life Sciences, Tsinghua University, Beijing, China
- State Key Laboratory of Chemical Oncogenomics, and Shenzhen Key Laboratory of Health Sciences and Technology, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, China
- Corresponding author
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18
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Obara K, Reynoso J, Hamada Y, Aoki Y, Kubota Y, Masaki N, Amoh Y, Hoffman RM. Hair follicle associated pluripotent (HAP) stem cells jump from transplanted whiskers to pelage follicles and stimulate hair growth. Sci Rep 2022; 12:21174. [PMID: 36476963 PMCID: PMC9729176 DOI: 10.1038/s41598-022-25383-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2022] [Accepted: 11/29/2022] [Indexed: 12/13/2022] Open
Abstract
Stimulation of hair growth in hair loss has been a difficult goal to achieve. Hair follicle-associated pluripotent (HAP) stem cells express nestin and have been shown to differentiate to multiple cell types including keratinocytes, neurons, beating cardiac muscles and numerous other cell types. HAP stem cells originate in the bulge area of the hair follicle and have been shown to migrate within and outside the hair follicle. In the present study, the upper part of vibrissa follicles from nestin-driven green-fluorescent protein (GFP) transgenic mice, containing GFP-expressing HAP stem cells, were transplanted in the dorsal area of athymic nude mice. Fluorescence microscopy and immunostaining showed the transplanted HAP stem cells jumped and targeted the bulge and hair bulb and other areas of the resident nude mouse pelage follicles where they differentiated to keratinocytes. These results indicate that transplanted nestin-GFP expressing HAP stem cells jumped from the upper part of the whisker follicles and targeted nude-mouse hair follicles, which are genetically deficient to grow normal hair shafts, and differentiated to keratinocytes to produce normal mature hair shafts. The resident nude-mouse pelage follicles targeted by jumping whisker HAP stem cells produced long hair shafts from numerous hair follicles for least 2 hair cycles during 36 days, demonstrations that HAP stem cells can stimulate hair growth. The present results for hair loss therapy are discussed.
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Affiliation(s)
- Koya Obara
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA.
- Department of Surgery, University of California San Diego, San Diego, CA, 92103, USA.
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, Kanagawa, 252-0374, Japan.
| | - Jose Reynoso
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, Kanagawa, 252-0374, Japan
| | - Yusuke Aoki
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA
- Department of Surgery, University of California San Diego, San Diego, CA, 92103, USA
| | - Yutaro Kubota
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA
- Department of Surgery, University of California San Diego, San Diego, CA, 92103, USA
| | - Noriyuki Masaki
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA
- Department of Surgery, University of California San Diego, San Diego, CA, 92103, USA
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, Kanagawa, 252-0374, Japan.
| | - Robert M Hoffman
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA.
- Department of Surgery, University of California San Diego, San Diego, CA, 92103, USA.
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19
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Emerging biomarker in carcinogenesis. Focus on Nestin. Postepy Dermatol Alergol 2022; 39:1001-1007. [PMID: 36686021 PMCID: PMC9837589 DOI: 10.5114/ada.2022.122599] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2021] [Accepted: 09/13/2021] [Indexed: 12/24/2022] Open
Abstract
Nestin is a protein belonging to class VI intermediate filaments, which is involved in organogenesis, cellular metabolism and cytoskeletal organisation. Originally found to be expressed in neuroepithelial stem cells, nestin is also expressed in other tissues. It plays an important role in the carcinogenesis and angiogenesis. Its increased expression in melanoma is associated with an aggressive course of the disease and poor prognosis. Research findings for non-melanocytic skin neoplasms are inconclusive. The aim of this paper was to systematize knowledge on the role of nestin in cancerogenesis. The authors focused in particular on the expression of nestin in skin malignancies, as well as on the potential role of nestin in the pathogenesis, prognosis and treatment of cutaneous neoplasms.
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20
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Yu T, Xu X, Mao H, Han X, Liu Y, Zhang H, Lai J, Gu J, Xia M, Hu C, Li D. Fenpropathrin exposure induces neurotoxicity in zebrafish embryos. FISH PHYSIOLOGY AND BIOCHEMISTRY 2022; 48:1539-1554. [PMID: 36266516 DOI: 10.1007/s10695-022-01134-9] [Citation(s) in RCA: 17] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/22/2022] [Accepted: 10/10/2022] [Indexed: 06/16/2023]
Abstract
Fenpropathrin has been a commonly used insecticide to control agricultural and household insects over a few decades. Up to now, fenpropathrin residue in soil and water has been often determined due to its widespread use, which poses serious threat to environment and aquatic organisms. The potential of fenpropathrin to affect aquatic lives is still poorly understood. In this study, we used zebrafish (Danio rerio) embryo as an experimental model system to evaluate the toxicity of fenpropathrin to the development of zebrafish nervous system. Zebrafish embryos were separately exposed to fenpropathrin at the dose of 0.016 mg/L, 0.032 mg/L, 0.064 mg/L, starting at 6 h post-fertilizationhpf (hpf) up to 96 hpf. The results showed that fenpropathrin exposure gives rise to physiological, behavioral, and neurodevelopmental impairments in zebrafish embryos, including enhanced acetylcholinesterase (AChE) activity, abnormal swimming behavior, karyopyknosis in brain cells, increased intercellular space, and uneven migration of neuron in brain area. In addition, the expressions of genes concerning neurodevelopment and neurotransmitter system were inhibited following fenpropathrin exposure. We also found that fenpropathrin exposure distinctly induced oxidative stress by increasing reactive oxygen species (ROS) generation and inhibiting the production of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). Expectedly, some apoptosis-associated genes were induced and the apoptosis appeared in the brain and heart cells of zebrafish embryos. Moreover, fenpropathrin exposure also inhibited the expressions of genes in Nrf2 signaling pathway, such as heme oxygenase-1 (HO-1) and SOD. In summary, the results of this study indicate that oxidative stress-triggered apoptosis may be an underlying fundamental of fenpropathrin-induced neurotoxicity in zebrafish embryos.
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Affiliation(s)
- Tingting Yu
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Xiaowen Xu
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Huiling Mao
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Xue Han
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Yulong Liu
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Hongying Zhang
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Jingli Lai
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Jianfeng Gu
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Mengling Xia
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Chengyu Hu
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China
| | - Dongming Li
- School of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China.
- School of Basic Medical Sciences, Fuzhou Medical College, Nanchang University, Fuzhou, 344000, Jiangxi, China.
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21
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Nestin is a marker of unipotent embryonic and adult progenitors differentiating into an epithelial cell lineage of the hair follicles. Sci Rep 2022; 12:17820. [PMID: 36280775 PMCID: PMC9592581 DOI: 10.1038/s41598-022-22427-2] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/25/2022] [Accepted: 10/14/2022] [Indexed: 01/19/2023] Open
Abstract
Nestin is an intermediate filament protein transiently expressed in neural stem/progenitor cells. We previously demonstrated that outer root sheath (ORS) keratinocytes of adult hair follicles (HFs) in mice descend from nestin-expressing cells, despite being an epithelial cell lineage. This study determined the exact stage when nestin-expressing ORS stem/precursor cells or their descendants appear during HF morphogenesis, and whether they are present in adult HFs. Using Nes-Cre/CAG-CAT-EGFP mice, in which enhanced green fluorescent protein (EGFP) is expressed following Cre-based recombination driven by the nestin promoter, we found that EGFP+ cells appeared in the epithelial layer of embryonic HFs as early as the peg stage. EGFP+ cells in hair pegs were positive for keratin 14 (K14) and K5, but not vimentin, SOX2, SOX10, or S100 alpha 6. Tracing of tamoxifen-induced EGFP+ cells in postnatal Nes-CreERT2/CAG-CAT-EGFP mice revealed labeling of some isthmus HF epithelial cells in the first anagen stage. EGFP+ cells in adult HFs were not immunolabeled for K15, an HF multipotent stem cell marker. However, when hairs were depilated in Nes-CreERT2/CAG-CAT-EGFP mice to induce the anagen stage after tamoxifen injection, the majority of ORS keratinocytes in depilation-induced anagen HFs were labeled for EGFP. Our findings indicate that nestin-expressing unipotent progenitor cells capable of differentiating into ORS keratinocytes are present in HF primordia and adult HFs.
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22
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Huang J, Deng R, Li W, Jiang M, Xiang AP, Zhang X. Nestin+ Mesenchymal Precursors Generate Distinct Spleen Stromal Cell Subsets and Have Immunomodulatory Function. Int J Mol Sci 2022; 23:ijms231911819. [PMID: 36233119 PMCID: PMC9569994 DOI: 10.3390/ijms231911819] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/02/2022] [Revised: 09/22/2022] [Accepted: 09/26/2022] [Indexed: 11/16/2022] Open
Abstract
Mesenchymal stromal cells (MSCs) are known to be widespread in many tissues and possess a broad spectrum of immunoregulatory properties. They have been used in the treatment of a variety of inflammatory diseases; however, the therapeutic effects are still inconsistent owing to their heterogeneity. Spleen stromal cells have evolved to regulate the immune response at many levels as they are bathed in a complex inflammatory milieu during infection. Therefore, it is unknown whether they have stronger immunomodulatory effects than their counterparts derived from other tissues. Here, using a transgenic mouse model expressing GFP driven by the Nestin (Nes) promoter, Nes-GFP+ cells from bone marrow and spleen were collected. Artificial lymphoid reconstruction in vivo was performed. Cell phenotype, inhibition of T cell inflammatory cytokines, and in vivo therapeutic effects were assessed. We observed Nes-GFP+ cells colocalized with splenic stromal cells and further demonstrated that these Nes-GFP+ cells had the ability to establish ectopic lymphoid-like structures in vivo. Moreover, we showed that the Nes-GFP+ cells possessed the characteristics of MSCs. Spleen-derived Nes-GFP+ cells exhibited greater immunomodulatory ability in vitro and more remarkable therapeutic efficacy in inflammatory diseases, especially inflammatory bowel disease (IBD) than bone marrow-derived Nes-GFP+ cells. Overall, our data showed that Nes-GFP+ cells contributed to subsets of spleen stromal populations and possessed the biological characteristics of MSCs with a stronger immunoregulatory function and therapeutic potential than bone marrow-derived Nes-GFP+ cells.
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Affiliation(s)
- Jing Huang
- Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China
| | - Ronghai Deng
- Department of Organ Transplantation, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China
| | - Weiqiang Li
- Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China
| | - Meihua Jiang
- Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China
| | - Andy Peng Xiang
- Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China
- Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China
- Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou 510080, China
| | - Xiaoran Zhang
- Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China
- Correspondence: ; Tel.: +86-20-87335982
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23
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Wang ZH, Liu LP, Zheng YW. Melanocyte stem cells in skin diseases and their potential in cell-based therapy. Histol Histopathol 2022; 37:937-953. [PMID: 35553404 DOI: 10.14670/hh-18-470] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 06/15/2023]
Abstract
Melanocytes have a complex function and play an important role in a variety of regulatory mechanisms in the human system. Melanocyte stem cells (MelSCs) serve as a reservoir to replenish the melanocytes by regenerating new ones, and they are capable of self-renewal and differentiation to maintain their homeostasis, repair, and regeneration in tissues. The numerical decrease and functional impairment of MelSCs may be closely related to the development and treatment response of many skin diseases. However, the current knowledge about MelSCs mainly comes from studies in mice, and little is known about human MelSC markers; especially, their markers are still unclear or lack consensus. This leads to uncertainty in clinical findings, which further limits our comprehensive understanding of pigmentary disorders and also hinders the progress of new treatments. Thus, in this review article, combined with our previous and current work, we summarize and update the recent advances in MelSC research, including the molecular markers of human MelSCs and their niche, as well as the association of MelSCs with skin diseases, including vitiligo, hair greying, and melanoma. Due to the limited tools available to explore the identified characteristics of human MelSCs, pluripotent stem cells can provide a new research model for further study, especially combined with CRISPR/Cas9 technology. The visualization of human MelSCs' development and differentiation can help to identify their molecular characteristics and understand their cellular fate dynamically, which will allow us not only to further explore their roles in associated diseases, but also to achieve MelSC-based cellular therapy.
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Affiliation(s)
- Zi-Han Wang
- Institute of Regenerative Medicine, and Department of Dermatology, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang, Jiangsu, China
| | - Li-Ping Liu
- Institute of Regenerative Medicine, and Department of Dermatology, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang, Jiangsu, China.
| | - Yun-Wen Zheng
- Institute of Regenerative Medicine, and Department of Dermatology, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang, Jiangsu, China
- Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, and School of Biotechnology and Health Sciences, Wuyi University, Jiangmen, Guangdong, China
- School of Medicine, Yokohama City University, Yokohama, Kanagawa, Japan
- Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan
- Department of Medical and Life Sciences, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda, Japan.
- Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, The University of Tokyo, Tokyo, Japan
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24
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Stüfchen I, Beckervordersandforth R, Fischer S, Kappelmann M, Bosserhoff AK, Beyer F. Two novel CreER T2 transgenic mouse lines to study melanocytic cells in vivo. Pigment Cell Melanoma Res 2022; 35:613-621. [PMID: 35920064 DOI: 10.1111/pcmr.13061] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2021] [Revised: 05/20/2022] [Accepted: 08/01/2022] [Indexed: 11/29/2022]
Abstract
The skin of adult mammals protects from radiation, physical and chemical insults. While melanocytes and melanocyte-producing stem cells contribute to proper skin function in healthy organisms, dysfunction of these cells can lead to the generation of malignant melanoma - the deadliest type of skin cancer. Addressing cells of the melanocyte lineage in vivo represents a prerequisite for the understanding of melanoma on cellular level and the development of preventive and treatment strategies. Here, the inducible Cre-loxP-system has emerged as a promising tool to specifically target, monitor and modulate cells in adult mice. Re-analysis of existing sequencing data sets of melanocytic cells revealed that genes with a known function in neural cells, including neural stem cells (Aldh1L1 and Nestin), are also expressed in melanocytic cells. Therefore, in this study we explored whether the promoter activity of Nestin and Aldh1L1 can serve to target cells of the melanocyte lineage using the inducible CreERT2 -loxP-system. Using an immunohistochemical approach and different time-points of analysis, we were able to map the melanocytic fate of recombined stem cells in the adult hair follicle of Nestin-CreERT2 and Aldh1L1-CreERT2 transgenic mice. Thus, we here present two new mouse models and propose their use to study and putatively modulate adult melanocytic cells in vivo.
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Affiliation(s)
- Isabel Stüfchen
- Institute of Biochemistry, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany
| | | | - Stefan Fischer
- Faculty of Computer Science, Deggendorf Institute of Technology, Deggendorf, Germany
| | - Melanie Kappelmann
- Faculty of Computer Science, Deggendorf Institute of Technology, Deggendorf, Germany
| | - Anja K Bosserhoff
- Institute of Biochemistry, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany
| | - Felix Beyer
- Institute of Biochemistry, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany
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25
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Wang Y, Wei T, Wang Q, Zhang C, Li K, Deng J. Resveratrol's neural protective effects for the injured embryoid body and cerebral organoid. BMC Pharmacol Toxicol 2022; 23:47. [PMID: 35820950 PMCID: PMC9275253 DOI: 10.1186/s40360-022-00593-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/20/2021] [Accepted: 07/06/2022] [Indexed: 11/10/2022] Open
Abstract
OBJECTIVE Resveratrol (RSV) is a polyphenol compound found in grapes, veratrum and other plants. It has been reported that RSV has anti-inflammatory, anti-oxidant, anti-cancer and other pharmacological effects. However, the impacts of RSV on development of nervous system are not understood well. The study aims to investigate RSV's neuroprotective effect during development and to provide a health care for pregnant women and their fetuses with RSV supplementation. METHODS In this study, we induced human induced pluripotent stem cells (hiPSCs) to form the embryoid bodies (EBs) and cerebral organoids (COs) with 3 dimensional (3D) culture. In the meantime, D-galactose (D-gal, 5 mg/ml) was used to make nervous injury model, and on the other hand, RSV with various doses, such as 2 μm/L, 10 μm/L, 50 μm/L, were applied to understand its neuroprotection. Therefore, the cultures were divided into control group, D-gal nervous injury group and RSV intervention groups. After that, the diameters of EBs and COs were measured regularly under a reverted microscope. In the meantime, the neural proliferation, cell apoptosis and the differentiation of germ layers were detected via immunofluorescence. RESULTS (1) D-gal could delay the development of EBs and COs; (2) RSV could rescue the atrophy of EBs and COs caused by D-gal; (3) RSV showed its neuroprotection, through promoting the neural cell proliferation, inhibiting apoptosis and accelerating the differentiation of germ layers. CONCLUSION RSV has a neuroprotective effect on the development of the nervous system, suggesting RSV supplementation may be necessary during the health care of pregnancy and childhood.
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Affiliation(s)
- Yanli Wang
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China
| | - Tingting Wei
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China
| | - Qiang Wang
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China
| | - Chaonan Zhang
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China
| | - Keyan Li
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China
| | - Jinbo Deng
- NHC Key Laboratory of Birth Defects Prevention, Henan Key Laboratory of Population Defects Prevention, Henan Institute of Reproduction Health Science and Technology, Zhengzhou, 450002, Henan Province, China.
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26
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Barnts K, Feng JQ, Qin C, Zhang H, Cheng YSL. Adenomatoid odontogenic tumor: evidence for a mixed odontogenic tumor. Oral Surg Oral Med Oral Pathol Oral Radiol 2022; 133:675-683. [PMID: 35165067 DOI: 10.1016/j.oooo.2021.11.005] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/15/2021] [Revised: 09/22/2021] [Accepted: 11/10/2021] [Indexed: 01/28/2023]
Abstract
OBJECTIVE Adenomatoid odontogenic tumor (AOT) was classified by the World Health Organization as a mixed odontogenic tumor in 1992 and reclassified without a clear rationale as an epithelium-only tumor in 2005. The purpose of this study was to investigate if there was any evidence to suggest AOT might be a mixed odontogenic tumor. STUDY DESIGN Immunohistochemical studies with nestin, dentin sialophosphoprotein (DSPP), cytokeratin, and vimentin were performed using 21 cases of AOT, and the staining results were analyzed according to the various morphologic patterns seen in AOT. Sirius red stain was used to detect the presence of collagen types I and III in AOT products. RESULTS Our results showed that 20 of 21 (95.23%), 0 of 21 (0%), 21 of 21 (100%), and 20 of 21 (95.23%) cases expressed nestin, DSPP, cytokeratin, and vimentin, respectively. Some cells in rosette/duct-like structures (RDSs) expressed nestin, vimentin, or both, without cytokeratin. Coexpression of vimentin and cytokeratin or of nestin, cytokeratin, and vimentin was noted in some cells. Sirius red staining was positive in eosinophilic products in RDSs, double-layered spheres, and dentinoids. CONCLUSION Although most AOT cells appear epithelial, there is a small population of cells expressing mesenchymal proteins and secreting collagen types I and III. This evidence suggests that AOT is a mixed odontogenic tumor.
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Affiliation(s)
- Kelcie Barnts
- Department of Oral and Maxillofacial Pathology, Medicine and Surgery, Kornberg School of Dentistry, Temple University, Philadelphia, USA
| | - Jian Q Feng
- Department of Biomedical Sciences, College of Dentistry, Texas A&M University, Dallas, Texas, USA
| | - Chunlin Qin
- Department of Biomedical Sciences, College of Dentistry, Texas A&M University, Dallas, Texas, USA
| | - Hua Zhang
- Department of Biomedical Sciences, College of Dentistry, Texas A&M University, Dallas, Texas, USA
| | - Yi-Shing Lisa Cheng
- Department of Diagnostic Sciences, College of Dentistry, Texas A&M University, Dallas, Texas, USA.
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Hair-follicle-associated pluripotent (HAP) stem cells differentiate into mature beating cardiomyocyte sheets on flexible substrates in vitro. Med Mol Morphol 2022; 55:248-257. [PMID: 35536435 DOI: 10.1007/s00795-022-00322-z] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/25/2022] [Accepted: 04/18/2022] [Indexed: 10/18/2022]
Abstract
Cardiomyocytes have been differentiated from various stem cells such as human embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC), but it is difficult to produce mature cardiomyocytes. We showed rat hair-follicle-associated pluripotent (HAP) stem cells have pluripotency and produced mature beating cardiomyocyte sheets differentiated from rat HAP stem cells. The upper parts of rat vibrissa hair follicles were cultured in 10% FBS DMEM and stained with antibodies of the ectoderm, mesoderm, endoderm system to show the differentiation of multiple cell types. Moreover, HAP stem cells were cultured under three different conditions to decide the most suitable culture conditions for making beating cardiomyocyte sheets. The beating cardiomyocyte sheets were shown to be mature by staining sarcomere structures. Isoproterenol alone and the combination of isoproterenol, activin A, bone morphogenetic protein 4 (BMP4) and basic fibroblast growth factor (bFGF) effectively induced beating long-fiber cardiomyocytes, which formed beating sheets, only in the presence of all four agents. Flexible substrates were essential for the differentiation of sheets of mature beating cardiomyocytes for HAP stem cells. The features of the cardiomyocytes differentiated from HAP stem cells demonstrate they have clinical potential for heart regeneration.
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28
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Erdoğan A, Mutlu HS, Solakoğlu S. Autologously transplanted dermis-derived cells alleviated monobenzone-induced vitiligo in mouse. Exp Dermatol 2022; 31:1355-1363. [PMID: 35538739 DOI: 10.1111/exd.14603] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/02/2022] [Revised: 04/26/2022] [Accepted: 05/08/2022] [Indexed: 12/01/2022]
Abstract
Vitiligo is a depigmentation disease which affects skin and hair follicles with a prevalence of 0.5-1% worldwide. In this study, we aimed to investigate treatmental potential of dermis-derived cells in monobenzone (MBEH)-induced mouse vitiligo model with light and electron microscopy. MBEH (40%) cream was topically applied to C57BL/6 mice until depigmentation occured in vitiligo and experimental groups. In experimental groups, dermis-derived cells obtained from back skin biopsy samples before induction of vitiligo, were injected intradermally to vitiligo mice. On day 3 and 15 after cell transplantation to experimental groups, skin biopsies were compared with biopsies of control and vitiligo groups. Dermis-derived cells obtained from back skin biopsy samples of experimental groups showed nestin and versican immunoreactivity. Melanin in hair follicles of control group was detected by histochemical stainings (Hematoxylin&Eosin and Fontana-Masson) whereas sparse melanin granules were observed in hair follicles of vitiligo group. In experimental groups, there was an increase in the number of hair follicles with melanin compared to vitiligo group. We observed MART-1 immunoreactive cells mostly around the hair follicles in control group and within dermis in vitiligo group. Electron microscopic investigation showed presence of melanosomes in hair follicles of control group and lacking in vitiligo group. In experimental groups, both type of hair follicles were observed with electron microscope. Our data suggest that autologously transplanted dermis-derived cells may be effective in vitiligo treatment by contrubuting to melanin production.
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Affiliation(s)
- Aslı Erdoğan
- İstanbul University, İstanbul Faculty of Medicine, Department of Histology and Embryology, İstanbul, Turkey.,İstanbul University, Graduate School of Health Sciences, İstanbul, Turkey.,İzmir Kȃtip Çelebi University, Faculty of Medicine, Department of Histology and Embryology, İzmir, Turkey
| | - Hasan Serdar Mutlu
- İstanbul University, İstanbul Faculty of Medicine, Department of Histology and Embryology, İstanbul, Turkey.,Giresun University, Faculty of Medicine, Department of Histology and Embryology, Giresun, Turkey
| | - Seyhun Solakoğlu
- İstanbul University, İstanbul Faculty of Medicine, Department of Histology and Embryology, İstanbul, Turkey
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Smurf2-induced degradation of SMAD2 causes inhibition of hair follicle stem cell differentiation. Cell Death Dis 2022; 8:160. [PMID: 35379779 PMCID: PMC8980066 DOI: 10.1038/s41420-022-00920-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2021] [Revised: 02/15/2022] [Accepted: 02/24/2022] [Indexed: 11/08/2022]
Abstract
Hair follicle stem cells (HFSCs) are implicated in the formation of hair follicles and epidermis. This study aims to clarify the role of SMAD2 in regulating the differentiation of HFSCs, which is involved with Smurf2. Functional assays were carried out in human HFSCs to assess the effect of SMAD2 and Smurf2 with altered expression on growth dynamics of HFSCs. Ubiquitination of SMAD2 and its protein stability were assessed. The binding relationship between NANOG and DNMT1 was assessed. A mouse skin wound model was induced to verify the effects of Smurf2/SMAD2/NANOG/DNMT1 on wound healing. SMAD2 overexpression was observed in HFSCs during differentiation and its ectopic expression contributed to promotion of differentiation and apoptosis of HFSCs while arresting cell proliferation. Mechanistic investigations indicated that Smurf2 promoted the ubiquitination and degradation of SMAD2, thus causing downregulation of SMAD2 expression. By this mechanism, NANOG expression was reduced and the subsequent DNMT1 transcriptional expression was also diminished, leading to suppression of differentiation and apoptosis of HFSCs while stimulating cell proliferation. Moreover, in vivo data showed that Smurf2 upregulation limited epidermal wound healing in mice by inhibiting the SMAD2/NANOG/DNMT1 axis. Our work proposed a potential target regarding SMAD2 restoration in promoting HFSC differentiation and skin wound healing.
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30
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Obara K, Shirai K, Hamada Y, Arakawa N, Yamane M, Takaoka N, Aki R, Hoffman RM, Amoh Y. Chronic spinal cord injury functionally repaired by direct implantation of encapsulated hair-follicle-associated pluripotent (HAP) stem cells in a mouse model: Potential for clinical regenerative medicine. PLoS One 2022; 17:e0262755. [PMID: 35085322 PMCID: PMC8794105 DOI: 10.1371/journal.pone.0262755] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/04/2021] [Accepted: 01/04/2022] [Indexed: 11/30/2022] Open
Abstract
Chronic spinal cord injury (SCI) is a highly debilitating and recalcitrant disease with limited treatment options. Although various stem cell types have shown some clinical efficacy for injury repair they have not for SCI. Hair-follicle-associated pluripotent (HAP) stem cells have been shown to differentiate into neurons, Schwan cells, beating cardiomyocytes and many other type of cells, and have effectively regenerated acute spinal cord injury in mouse models. In the present report, HAP stem cells from C57BL/6J mice, encapsulated in polyvinylidene fluoride membranes (PFM), were implanted into the severed thoracic spinal cord of C57BL/6J or athymic nude mice in the early chronic phase. After implantation, HAP stem cells differentiated to neurons, astrocytes and oligodendrocytes in the regenerated thoracic spinal cord of C57BL/6J and nude mice. Quantitative motor function analysis, with the Basso Mouse Scale for Locomotion (BMS) score, demonstrated a significant functional improvement in the HAP-stem-cell-implanted mice, compared to non-implanted mice. HAP stem cells have critical advantages over other stem cells: they do not develop teratomas; do not loose differentiation ability when cryopreserved and thus are bankable; are autologous, readily obtained from anyone; and do not require genetic manipulation. HAP stem cells therefore have greater clinical potential for SCI repair than induced pluripotent stem cells (iPSCs), neuronal stem cells (NSCs)/neural progenitor cells (NPCs) or embryonic stem cells (ESCs). The present report demonstrates future clinical potential of HAP-stem-cell repair of chronic spinal cord injury, currently a recalcitrant disease.
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Affiliation(s)
- Koya Obara
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Kyoumi Shirai
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nobuko Arakawa
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Michiko Yamane
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Nanako Takaoka
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Ryoichi Aki
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
| | - Robert M. Hoffman
- AntiCancer, Inc., San Diego, California, United States of America
- Department of Surgery, University of California San Diego, San Diego, California, United States of America
- * E-mail: (YA); (RMH)
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
- * E-mail: (YA); (RMH)
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31
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Singh S, Muthuvel K. Role of Hair Transplantation in Scarring Alopecia-To Do or Not to Do. Indian J Plast Surg 2022; 54:501-506. [PMID: 34984092 PMCID: PMC8719951 DOI: 10.1055/s-0041-1739246] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022] Open
Abstract
Alopecia in the scalp region leads to psychosocial embarrassment for an individual. Alopecia could be due to several reasons, including genetic, hormonal, traumatic and infections. Cicatricial alopecias (CAs) are considered as trichological emergency, since their progression is rapid and always results in permanent hair loss. The pathogenesis, disease progression and prognosis of CA are poorly understood, and the treatment process is still evolving. An early diagnosis must be established, and aggressive treatment protocol should be followed in the management of scarring alopecia. This article presents various aspects of CA and determines whether hair transplant (HT) should be done in this condition.
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Affiliation(s)
- Sukhbir Singh
- Department of Plastic Surgery, Resplendent the Cosmetic Studio, New Delhi, India
| | - Kumaresan Muthuvel
- Department of Dermatology, Cutis Skin Clinic and Hair Transplant Centre, Coimbatore, Tamil Nadu, India
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32
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Hejazian LB, Akbarnejad Z, Moghani Ghoroghi F, Esmaeilzade B, Chaibakhsh S. Augmenting Peripheral Nerve Regeneration Using Hair Follicle Stem Cells in Rats. Basic Clin Neurosci 2022; 13:57-70. [PMID: 36589026 PMCID: PMC9790101 DOI: 10.32598/bcn.2021.2240.1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2019] [Revised: 02/23/2020] [Accepted: 08/01/2020] [Indexed: 01/04/2023] Open
Abstract
Introduction Cell therapy is the most advanced treatment of peripheral nerve injury. This study aimed to determine the effects of transplantation of hair follicle stem cells on the regeneration of the sciatic nerve injury in rats. Methods The bulge region of the rat whisker were isolated and cultured. Morphological and biological features of the cultured bulge cells were observed by light microscopy and immunocytochemistry methods. Percentages of CD34, K15, and nestin cell markers expression were demonstrated by flow cytometry. Rats were randomly divided into 3 groups of injury, epineurium, and epineurium with cells in which rat Hair Follicular Stem Cells (rHFSCs) were injected into the site of the nerve cut. HFSCs were labeled with Bromodeoxyuridine (BrdU), and double-labeling immunofluorescence was performed to study the survival and differentiation of the grafted cells. After 8 weeks, electrophysiological, histological, and immunocytochemical analysis assessments were performed. Results Rat hair follicle stem cells are suitable for cell culture, proliferation, and differentiation. The results suggest that transplantation of rat hair follicle stem cells can regenerate sciatic nerve injury; moreover, electrophysiology and histology examinations show that sciatic nerve repair was more effective in the epineurium with cell group than in the other experimental group (P<0.05). Conclusion The achieved results propose that hair follicle stem cells improve axonal growth and functional recovery after peripheral nerve injury. Highlights This study showed that rat hair follicle stem cells are suitable for cell culture, proliferation and differentiationThe results suggested that transplantation of rat hair follicle stem cells had the potential capability of regenerating sciatic nerve injuryEvidence of electrophysiology and histology showed Concomitant use of epineurium with hair follicle stem cell was more effective repairment. Plain Language Summary Although repairing damaged peripheral nerves has always been a medical challenge, but peripheral nerve injury has been successfully repaired using various procedures such as nerve auto-graft or stem cell therapy. The functional reconstruction is the most important after therapy because of that primary nerve repair or use of nerve autograft, are still accepted as golden standard methods for treatment. Considerable recent interest has been focused on adult stem cells for both research and clinical applications. A highly promising source of relatively abundant and accessible, active, multipotent adult stem cells are obtained from hair follicles. In research the hair follicle stem cells implanted into the gap region of a severed sciatic nerve injury greatly enhanced the rate of nerve regeneration and the restoration of nerve function. Time is one of the several aspects require specific attention in the clinical treatment of peripheral nerve injury. Because delay of nerve injury treatment may cause neurobiological alterations in neurons and Schwann cells, impairing nerve functional recovery and affect neuron survival. In this study, concluded that stem cell injection 2 weeks after injury in the damaged nerve epineurium repairs nerve fibers, while electrophysiology of the leg muscles showed that muscle function was significantly improved. It indicates the repair of muscular innervation and nerve repair. The results pave the way for further research on this topic.
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Affiliation(s)
- Leila Beigom Hejazian
- Department of Anatomy, School of Medicine, Babol University of Medical Sciences, Babol, Iran
| | - Zeinab Akbarnejad
- ENT and Head & Neck Research Center, The five Senses Health Institute, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | | | - Banafshe Esmaeilzade
- Department of Anatomy, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran
| | - Samira Chaibakhsh
- Eye Research Center, the Five Senses Institute, Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran
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Abreu CM, Marques AP. Recreation of a hair follicle regenerative microenvironment: Successes and pitfalls. Bioeng Transl Med 2022; 7:e10235. [PMID: 35079623 PMCID: PMC8780054 DOI: 10.1002/btm2.10235] [Citation(s) in RCA: 21] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/21/2021] [Revised: 05/15/2021] [Accepted: 05/18/2021] [Indexed: 12/19/2022] Open
Abstract
The hair follicle (HF) is an exquisite skin appendage endowed with cyclical regenerative capacity; however, de novo follicle formation does not naturally occur. Consequently, patients suffering from extensive skin damage or hair loss are deprived of the HF critical physiological and/or aesthetic functions, severally compromising skin function and the individual's psychosocial well-being. Translation of regenerative strategies has been prevented by the loss of trichogenic capacity that relevant cell populations undergo in culture and by the lack of suitable human-based in vitro testing platforms. Here, we provide a comprehensive overview of the major difficulties associated with HF regeneration and the approaches used to overcome these drawbacks. We describe key cellular requirements and discuss the importance of the HF extracellular matrix and associated signaling for HF regeneration. Finally, we summarize the strategies proposed so far to bioengineer human HF or hair-bearing skin models and disclose future trends for the field.
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Affiliation(s)
- Carla M. Abreu
- 3B's Research Group, I3Bs ‐ Research Institute on Biomaterials, Biodegradables and Biomimetics, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineAvePark–Parque de Ciência e Tecnologia, University of MinhoGuimarãesPortugal
- ICVS/3B's–PT Government Associate LaboratoryGuimarãesPortugal
| | - Alexandra P. Marques
- 3B's Research Group, I3Bs ‐ Research Institute on Biomaterials, Biodegradables and Biomimetics, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative MedicineAvePark–Parque de Ciência e Tecnologia, University of MinhoGuimarãesPortugal
- ICVS/3B's–PT Government Associate LaboratoryGuimarãesPortugal
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Akbari S, Hooshmandi E, Bayat M, Borhani Haghighi A, Salehi MS, Pandamooz S, Yousefi Nejad A, Haghani M. The neuroprotective properties and therapeutic potential of epidermal neural crest stem cells transplantation in a rat model of vascular dementia. Brain Res 2021; 1776:147750. [PMID: 34896332 DOI: 10.1016/j.brainres.2021.147750] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/06/2021] [Revised: 11/17/2021] [Accepted: 12/06/2021] [Indexed: 01/04/2023]
Abstract
INTRODUCTION The incidence rate of senile dementia is rising, and there is no definite cure for it yet. Cell therapy, as a new investigational approach, has shown promising results. Hair bulges with abundant easily accessible neural stem cells permit autologous implantation in irreversible neurodegenerative disorders. METHODS Fifty rats were randomly divided into 5 groups of control, sham-operation, two-common carotid vessel-occlusion rats that received vehicle (2VO + V), 2VO rats that received 1 × 106 epidermal stem cells (2VO + ESC1), and 2VO rats that received 2.5 × 106 epidermal stem cells (2VO + ESC2) in 300 µl PBS intravenously on days 4, 9, and 14 after surgery. The epidermal neural crest stem cells (EPI-NCSCs) were isolated from hair follicles of rat whiskers. The open-field, passive avoidance, and Morris water maze were used as behavioral tests. The basal-synaptic transmission, long-term potentiation (LTP), and short-term synaptic plasticity were evaluated by field-potential recording of the CA1 hippocampal area. RESULTS 30 days after the first transplantation in the 2VO + ESC1 group, functional recovery was prominent in anxiety and fear memory compared to the 2VO + ESC2 group, while LTP induction was recovered in both groups of grafted animals without improvement in basal synaptic transmission. These positive recoveries may be related to the release of different neurotrophic factors from grafted cells that can stimulate endogenous neurogenesis and synaptic plasticity. CONCLUSIONS Our results showed that EPI-NCSCs implantation could rescue LTP and cognitive disability in 2VO rats, while transplantation of 1 million cells showed better performance relative to 2.5 million cells.
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Affiliation(s)
- Somayeh Akbari
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Etrat Hooshmandi
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Mahnaz Bayat
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
| | | | - Mohammad Saied Salehi
- Clinical Neurology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Sareh Pandamooz
- Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Amirhossein Yousefi Nejad
- Department of Veterinary Medicine, Faculty of Veterinary Medicine. Islamic Azad University of Kazeroon, Shiraz, Iran
| | - Masoud Haghani
- Department of Physiology, The Medical School, Shiraz University of Medical Sciences, Shiraz, Iran; Histomorphometry and Stereology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran.
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35
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Peterson A, Nair L. Hair Follicle Stem Cells for Tissue Regeneration. TISSUE ENGINEERING PART B-REVIEWS 2021; 28:695-706. [PMID: 34238037 PMCID: PMC9419938 DOI: 10.1089/ten.teb.2021.0098] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
With the positive outcomes of various cell therapies currently under pre-clinical and clinical studies, there is a significant interest in novel stem cell sources with unique therapeutic properties. Studies over the past two decades or so demonstrated the feasibility to isolate multipotent/pluripotent stem cells from hair follicles. The easy accessibility, high proliferation and differentiation ability as well as lack of ethical concerns associated with this stem cell source make hair follicle stem cells (HFSCs) attractive candidate for cell therapy and tissue engineering. This review discusses the various stem cell types identified in rodent and human hair follicles and ongoing studies on the potential use of HFSCs for skin, bone, cardio-vascular, and nerve tissue engineering.
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Affiliation(s)
- Alyssa Peterson
- University of Connecticut, 7712, Storrs, Connecticut, United States;
| | - Lakshmi Nair
- University of Connecticut Health Center, 21654, Orthopaedic Surgery, Farmington, Connecticut, United States;
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Characterisation of Neurospheres-Derived Cells from Human Olfactory Epithelium. Cells 2021; 10:cells10071690. [PMID: 34359860 PMCID: PMC8307784 DOI: 10.3390/cells10071690] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2021] [Revised: 06/25/2021] [Accepted: 06/30/2021] [Indexed: 11/17/2022] Open
Abstract
A major problem in psychiatric research is a deficit of relevant cell material of neuronal origin, especially in large quantities from living individuals. One of the promising options is cells from the olfactory neuroepithelium, which contains neuronal progenitors that ensure the regeneration of olfactory receptors. These cells are easy to obtain with nasal biopsies and it is possible to grow and cultivate them in vitro. In this work, we used RNAseq expression profiling and immunofluorescence microscopy to characterise neurospheres-derived cells (NDC), that simply and reliably grow from neurospheres (NS) obtained from nasal biopsies. We utilized differential expression analysis to explore the molecular changes that occur during transition from NS to NDC. We found that processes associated with neuronal and vascular cells are downregulated in NDC. A comparison with public transcriptomes revealed a depletion of neuronal and glial components in NDC. We also discovered that NDC have several metabolic features specific to neuronal progenitors treated with the fungicide maneb. Thus, while NDC retain some neuronal/glial identity, additional protocol alterations are needed to use NDC for mass sample collection in psychiatric research.
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Yuan H, Hu H, Chen R, Mu W, Wang L, Li Y, Chen Y, Ding X, Xi Y, Mao S, Jiang M, Chen J, He Y, Wang L, Dong Y, Tou J, Chen W. Premigratory neural crest stem cells generate enteric neurons populating the mouse colon and regulating peristalsis in tissue-engineered intestine. Stem Cells Transl Med 2021; 10:922-938. [PMID: 33481357 PMCID: PMC8133337 DOI: 10.1002/sctm.20-0469] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2020] [Revised: 12/26/2020] [Accepted: 01/03/2021] [Indexed: 12/13/2022] Open
Abstract
Hirschsprung's disease (HSCR) is a common congenital defect. It occurs when bowel colonization by neural crest-derived enteric nervous system (ENS) precursors is incomplete during the first trimester of pregnancy. Several sources of candidate cells have been previously studied for their capacity to regenerate the ENS, including enteric neural crest stem cells (En-NCSCs) derived from native intestine or those simulated from human pluripotent stem cells (hPSCs). However, it is not yet known whether the native NCSCs other than En-NCSCs would have the potential of regenerating functional enteric neurons and producing neuron dependent motility under the intestinal environment. The present study was designed to determine whether premigratory NCSCs (pNCSCs), as a type of the nonenteric NCSCs, could form enteric neurons and mediate the motility. pNCSCs were firstly transplanted into the colon of adult mice, and were found to survive, migrate, differentiate into enteric neurons, and successfully integrate into the adult mouse colon. When the mixture of pNCSCs and human intestinal organoids was implanted into the subrenal capsule of nude mice and grown into the mature tissue-engineered intestine (TEI), the pNCSCs-derived neurons mediated neuron-dependent peristalsis of TEI. These results show that the pNCSCs that were previously assumed to not be induced by intestinal environment or cues can innervate the intestine and establish neuron-dependent motility. Future cell candidates for ENS regeneration may include nonenteric NCSCs.
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Affiliation(s)
- Huipu Yuan
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Institute of Translational Medicine, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Hui Hu
- Department of Laboratory MedicineHangzhou Medical CollegeHangzhouPeople's Republic of China
| | - Rui Chen
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Department of Neonatal SurgeryChildren's Hospital, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Wenbo Mu
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Institute of Translational Medicine, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Liangliang Wang
- Interdisciplinary Institutes of Neuroscience and Technology, Qiushi Academy for Advanced Studies, Zhejiang UniversityHangzhouPeople's Republic of China
| | - Ying Li
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Institute of Translational Medicine, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Yuelei Chen
- Cell Bank/Stem Cell BankInstitute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesShanghaiPeople's Republic of China
| | - Xiaoyan Ding
- Cell Bank/Stem Cell BankInstitute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesShanghaiPeople's Republic of China
| | - Yongmei Xi
- Institute of Genetics and Department of Genetics, Division of Human Reproduction and Developmental Genetics of the Women's HospitalZhejiang University School of MedicineHangzhouPeople's Republic of China
| | - ShanShan Mao
- Department of Internal NeurologyChildren's Hospital, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Mizu Jiang
- Department of GastroenterologyZhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Jie Chen
- Department of Pediatric SurgeryXinhua Hospital, Shanghai Jiao Tong University School of MedicineShanghaiPeople's Republic of China
| | - Yong He
- State Key Laboratory of Fluid Power and Mechatronic Systems School of Mechanical EngineeringZhejiang UniversityHangzhouPeople's Republic of China
| | - Lang Wang
- Interdisciplinary Institutes of Neuroscience and Technology, Qiushi Academy for Advanced Studies, Zhejiang UniversityHangzhouPeople's Republic of China
| | - Yi Dong
- Key Laboratory of Adolescent Health Assessment and Exercise Intervention of Ministry of Education, School of Physical Education & Health CareEast China Normal UniversityShanghaiPeople's Republic of China
| | - Jinfa Tou
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Department of Neonatal SurgeryChildren's Hospital, Zhejiang University School of MedicineHangzhouPeople's Republic of China
| | - Wei Chen
- Institute of Translational Medicine, and Children's Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child HealthHangzhouPeople's Republic of China
- Institute of Translational Medicine, Zhejiang University School of MedicineHangzhouPeople's Republic of China
- Department of NeurobiologyInstitute of Neuroscience, NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University School of MedicineHangzhouPeople's Republic of China
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Zhai X, Gong M, Peng Y, Yang D. Effects of UV Induced-Photoaging on the Hair Follicle Cycle of C57BL6/J Mice. Clin Cosmet Investig Dermatol 2021; 14:527-539. [PMID: 34040410 PMCID: PMC8140904 DOI: 10.2147/ccid.s310487] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2021] [Accepted: 05/05/2021] [Indexed: 12/16/2022]
Abstract
Purpose To study the changes in the hair follicle cycle and related stem cells induced by photoaging to establish a mouse model of senescence in hair follicles. Methods There were 54 C57BL6/J mice randomly divided into three groups. The UVA group and the UVB group underwent photoaging induced by UV lamps for 8 weeks. Changes in skin and the hair follicle cycle were compared by physical signs, dermoscopy, and hematoxylin and eosin and Masson's staining in each group. Western blot, immunohistochemistry, and RT-qPCR were carried out to test canonical proteins and gene expression of the Wnt signaling pathway in the samples. Immunofluorescence was chosen to show variations in the stem cells related to the hair follicle cycle. Results There were more gray hairs in the UVA group than the other groups (P<0.05). Both diameter of the hair shaft and depth of hair root were significantly decreased in the UV groups (P<0.05). Stem cells and melanocytes of the hair follicles were reduced in the UVA group. UV, especially UVB, up-regulated the expression of the Wnt signaling pathway and prolonged anagen and telogen phases in the hair follicles, compared with the control group (P<0.05). Conclusion By decreasing the number of stem cells related to hair follicles, UVA induces hair follicle photoaging characterized by hair follicle miniaturization and gray hairs. UV up-regulated the expression of the Wnt signaling pathway, and the hair follicle cycle was significantly prolonged by UVB.
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Affiliation(s)
- Xu Zhai
- Department of Plastic Surgery, The 2nd Hospital of Harbin Medical University, Harbin, People's Republic of China
| | - Meihua Gong
- Department of Plastic Surgery, The 2nd Hospital of Harbin Medical University, Harbin, People's Republic of China.,Department of Plastic and Cosmetic Surgery, Shenzhen People's Hospital, Second Affiliated Hospital of Jinan University Medical College, Shenzhen, People's Republic of China
| | - Yixuan Peng
- Department of Plastic Surgery, The 2nd Hospital of Harbin Medical University, Harbin, People's Republic of China
| | - Daping Yang
- Department of Plastic Surgery, The 2nd Hospital of Harbin Medical University, Harbin, People's Republic of China
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Li H, Masieri FF, Schneider M, Bartella A, Gaus S, Hahnel S, Zimmerer R, Sack U, Maksimovic-Ivanic D, Mijatovic S, Simon JC, Lethaus B, Savkovic V. The Middle Part of the Plucked Hair Follicle Outer Root Sheath Is Identified as an Area Rich in Lineage-Specific Stem Cell Markers. Biomolecules 2021; 11:biom11020154. [PMID: 33503918 PMCID: PMC7912066 DOI: 10.3390/biom11020154] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2020] [Revised: 01/15/2021] [Accepted: 01/19/2021] [Indexed: 12/13/2022] Open
Abstract
Hair follicle outer root sheath (ORS) is a putative source of stem cells with therapeutic capacity. ORS contains several multipotent stem cell populations, primarily in the distal compartment of the bulge region. However, the bulge is routinely obtained using invasive isolation methods, which require human scalp tissue ex vivo. Non-invasive sampling has been standardized by means of the plucking procedure, enabling to reproducibly obtain the mid-ORS part. The mid-ORS shows potential for giving rise to multiple stem cell populations in vitro. To demonstrate the phenotypic features of distal, middle, and proximal ORS parts, gene and protein expression profiles were studied in physically separated portions. The mid-part of the ORS showed a comparable or higher NGFR, nestin/NES, CD34, CD73, CD44, CD133, CK5, PAX3, MITF, and PMEL expression on both protein and gene levels, when compared to the distal ORS part. Distinct subpopulations of cells exhibiting small and round morphology were characterized with flow cytometry as simultaneously expressing CD73/CD271, CD49f/CD105, nestin, and not CK10. Potentially, these distinct subpopulations can give rise to cultured neuroectodermal and mesenchymal stem cell populations in vitro. In conclusion, the mid part of the ORS holds the potential for yielding multiple stem cells, in particular mesenchymal stem cells.
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Affiliation(s)
- Hanluo Li
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
| | | | - Marie Schneider
- Clinic for Hematology, Cell Therapy and Hemostaseology, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Alexander Bartella
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
| | - Sebastian Gaus
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
| | - Sebastian Hahnel
- Polyclinic for Dental Prosthetics and Material Sciences, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Rüdiger Zimmerer
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
| | - Ulrich Sack
- Medical Faculty, Institute for Clinical Immunology, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Danijela Maksimovic-Ivanic
- Department of Immunology, Institute for Biological Research ‘Sinisa Stankovic’ (IBISS)-National Institute of Republic of Serbia, University of Belgrade, 11000 Belgrade, Serbia; (D.M.-I.); (S.M.)
| | - Sanja Mijatovic
- Department of Immunology, Institute for Biological Research ‘Sinisa Stankovic’ (IBISS)-National Institute of Republic of Serbia, University of Belgrade, 11000 Belgrade, Serbia; (D.M.-I.); (S.M.)
| | - Jan-Christoph Simon
- Clinic for Dermatology, Venereology and Allergology, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Bernd Lethaus
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
| | - Vuk Savkovic
- Department of Cranial Maxillofacial Plastic Surgery, University Hospital Leipzig, 04103 Leipzig, Germany; (H.L.); (A.B.); (S.G.); (R.Z.); (B.L.)
- Correspondence: ; Tel.: +49-341-9721115
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Wang HY, Li SW, Wu TH, Wu ZH, Guo JX. The effect of androgen on wool follicles and keratin production in Hetian sheep. BRAZ J BIOL 2021; 81:526-536. [PMID: 33470295 DOI: 10.1590/1519-6984.224056] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2019] [Accepted: 03/18/2020] [Indexed: 12/14/2022] Open
Abstract
To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.
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Affiliation(s)
- H Y Wang
- Tarim University, College of Life Sciences, Key Laboratory of Protection e Utilization of Biological Resources in Tarim Basin, Alar, Xinjiang, China
| | - S W Li
- Tarim University, College of Life Sciences, Key Laboratory of Protection e Utilization of Biological Resources in Tarim Basin, Alar, Xinjiang, China
| | - T H Wu
- Tarim University, College of Life Sciences, Key Laboratory of Protection & Utilization of Biological Resources in Tarim Basin, Alar, Xinjiang, China
| | - Z H Wu
- Tarim University, College of Life Sciences, Key Laboratory of Protection & Utilization of Biological Resources in Tarim Basin, Alar, Xinjiang, China
| | - J X Guo
- Tarim University, College of Life Sciences, Key Laboratory of Protection & Utilization of Biological Resources in Tarim Basin, Alar, Xinjiang, China
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Tenorio-Mina A, Cortés D, Esquivel-Estudillo J, López-Ornelas A, Cabrera-Wrooman A, Lara-Rodarte R, Escobedo-Avila I, Vargas-Romero F, Toledo-Hernández D, Estudillo E, Acevedo-Fernández JJ, Tapia JSO, Velasco I. Human Keratinocytes Adopt Neuronal Fates After In Utero Transplantation in the Developing Rat Brain. Cell Transplant 2021; 30:963689720978219. [PMID: 33435710 PMCID: PMC7809298 DOI: 10.1177/0963689720978219] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2019] [Revised: 11/02/2020] [Accepted: 11/12/2020] [Indexed: 11/30/2022] Open
Abstract
Human skin contains keratinocytes in the epidermis. Such cells share their ectodermal origin with the central nervous system (CNS). Recent studies have demonstrated that terminally differentiated somatic cells can adopt a pluripotent state, or can directly convert its phenotype to neurons, after ectopic expression of transcription factors. In this article we tested the hypothesis that human keratinocytes can adopt neural fates after culturing them in suspension with a neural medium. Initially, keratinocytes expressed Keratins and Vimentin. After neural induction, transcriptional upregulation of NESTIN, SOX2, VIMENTIN, SOX1, and MUSASHI1 was observed, concomitant with significant increases in NESTIN detected by immunostaining. However, in vitro differentiation did not yield the expression of neuronal or astrocytic markers. We tested the differentiation potential of control and neural-induced keratinocytes by grafting them in the developing CNS of rats, through ultrasound-guided injection. For this purpose, keratinocytes were transduced with lentivirus that contained the coding sequence of green fluorescent protein. Cell sorting was employed to select cells with high fluorescence. Unexpectedly, 4 days after grafting these cells in the ventricles, both control and neural-induced cells expressed green fluorescent protein together with the neuronal proteins βIII-Tubulin and Microtubule-Associated Protein 2. These results support the notion that in vivo environment provides appropriate signals to evaluate the neuronal differentiation potential of keratinocytes or other non-neural cell populations.
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Affiliation(s)
- Andrea Tenorio-Mina
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
| | - Daniel Cortés
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
| | - Joel Esquivel-Estudillo
- Facultad de Medicina, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
- Unidad de Diagnóstico y Medicina Molecular, “Dr. Ruy Pérez Tamayo”, Hospital del Niño Morelense/Facultad de Medicina-UAEM, Zapata, Morelos, Mexico
| | - Adolfo López-Ornelas
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
- División de Investigación, Hospital Juárez de México, Mexico City, Mexico
| | - Alejandro Cabrera-Wrooman
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Unidad de Diagnóstico y Medicina Molecular, “Dr. Ruy Pérez Tamayo”, Hospital del Niño Morelense/Facultad de Medicina-UAEM, Zapata, Morelos, Mexico
- Instituto Nacional de Rehabilitación, Mexico City, Mexico
| | - Rolando Lara-Rodarte
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
| | - Itzel Escobedo-Avila
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
| | - Fernanda Vargas-Romero
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
| | - Diana Toledo-Hernández
- Unidad de Diagnóstico y Medicina Molecular, “Dr. Ruy Pérez Tamayo”, Hospital del Niño Morelense/Facultad de Medicina-UAEM, Zapata, Morelos, Mexico
- Centro de Investigación en Dinámica Celular, Instituto de Ciencias, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
| | - Enrique Estudillo
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
| | | | - Jesús Santa-Olalla Tapia
- Facultad de Medicina, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
- Unidad de Diagnóstico y Medicina Molecular, “Dr. Ruy Pérez Tamayo”, Hospital del Niño Morelense/Facultad de Medicina-UAEM, Zapata, Morelos, Mexico
| | - Iván Velasco
- Instituto de Fisiología Celular - Neurociencias, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico
- Laboratorio de Reprogramación Celular, Instituto Nacional de Neurología y Neurocirugía “Manuel Velasco Suárez”, Mexico City, Mexico
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Zhang X, Tang H, Mao S, Li B, Zhou Y, Yue H, Wang D, Wang Y, Fu J. Transplanted hair follicle stem cells migrate to the penumbra and express neural markers in a rat model of cerebral ischaemia/reperfusion. Stem Cell Res Ther 2020; 11:413. [PMID: 32967732 PMCID: PMC7510278 DOI: 10.1186/s13287-020-01927-w] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2020] [Revised: 08/16/2020] [Accepted: 09/07/2020] [Indexed: 02/08/2023] Open
Abstract
BACKGROUND Ischaemic stroke has become the main cause of death and severe neurological disorders, for which effective restorative treatments are currently limited. While stem cell transplantation offers therapeutic potential through neural regeneration, this approach is associated with the challenges of limited applicable sources. Hair follicle stem cells (HFSCs) are multipotential cells that can differentiate into ectodermal and mesodermal lineages and proliferate for long periods. The therapeutic potentials of HFSCs have not been investigated in ischaemic stroke models, and therefore, in this study, we aimed to determine whether they could survive and migrate to ischaemic areas after a stroke attack. METHODS A rat model of middle cerebral artery ischaemia/reperfusion was established and intravenously administered HFSCs. The potential of HFSCs to migrate and differentiate into neuron-like cells as well as their ability to reduce the infarct size was evaluated. Rat brain tissue samples were collected 2 weeks after cell transplantation and analysed via TTC staining, immunofluorescence and immunohistochemistry methods. The data were statistically analysed and presented as the means ± standard deviations. RESULTS Intravenously administrated rat HFSCs were able to migrate to the penumbra where they expressed neuron-specific markers, reduced the infarct volume and promoted neurological recovery. CONCLUSION HFSC transplantation has therapeutic potential for ischaemic stroke and is, therefore, worthy of further investigation toward possible clinical development for treating stroke patients.
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Affiliation(s)
- Xuemei Zhang
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Hao Tang
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Senlin Mao
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Bing Li
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Yinglian Zhou
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Hui Yue
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Duo Wang
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Yifei Wang
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China
| | - Jin Fu
- Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No.246 Xuefu Road, Nangang District, Harbin, 150086, Heilongjiang Province, China.
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O'Sullivan JDB, Nicu C, Picard M, Chéret J, Bedogni B, Tobin DJ, Paus R. The biology of human hair greying. Biol Rev Camb Philos Soc 2020; 96:107-128. [PMID: 32965076 DOI: 10.1111/brv.12648] [Citation(s) in RCA: 46] [Impact Index Per Article: 9.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/02/2020] [Revised: 08/17/2020] [Accepted: 08/20/2020] [Indexed: 12/12/2022]
Abstract
Hair greying (canities) is one of the earliest, most visible ageing-associated phenomena, whose modulation by genetic, psychoemotional, oxidative, senescence-associated, metabolic and nutritional factors has long attracted skin biologists, dermatologists, and industry. Greying is of profound psychological and commercial relevance in increasingly ageing populations. In addition, the onset and perpetuation of defective melanin production in the human anagen hair follicle pigmentary unit (HFPU) provides a superb model for interrogating the molecular mechanisms of ageing in a complex human mini-organ, and greying-associated defects in bulge melanocyte stem cells (MSCs) represent an intriguing system of neural crest-derived stem cell senescence. Here, we emphasize that human greying invariably begins with the gradual decline in melanogenesis, including reduced tyrosinase activity, defective melanosome transfer and apoptosis of HFPU melanocytes, and is thus a primary event of the anagen hair bulb, not the bulge. Eventually, the bulge MSC pool becomes depleted as well, at which stage greying becomes largely irreversible. There is still no universally accepted model of human hair greying, and the extent of genetic contributions to greying remains unclear. However, oxidative damage likely is a crucial driver of greying via its disruption of HFPU melanocyte survival, MSC maintenance, and of the enzymatic apparatus of melanogenesis itself. While neuroendocrine factors [e.g. alpha melanocyte-stimulating hormone (α-MSH), adrenocorticotropic hormone (ACTH), ß-endorphin, corticotropin-releasing hormone (CRH), thyrotropin-releasing hormone (TRH)], and micropthalmia-associated transcription factor (MITF) are well-known regulators of human hair follicle melanocytes and melanogenesis, how exactly these and other factors [e.g. thyroid hormones, hepatocyte growth factor (HGF), P-cadherin, peripheral clock activity] modulate greying requires more detailed study. Other important open questions include how HFPU melanocytes age intrinsically, how psychoemotional stress impacts this process, and how current insights into the gerontobiology of the human HFPU can best be translated into retardation or reversal of greying.
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Affiliation(s)
- James D B O'Sullivan
- Dr. Philip Frost Department for Dermatology and Cutaneous Surgery, University of Miami, Miami, Florida, 33136, U.S.A
| | - Carina Nicu
- Dr. Philip Frost Department for Dermatology and Cutaneous Surgery, University of Miami, Miami, Florida, 33136, U.S.A
| | - Martin Picard
- Departments of Psychiatry and Neurology, Columbia University Irving Medical Center, 622 W 168th Street, PH1540N, New York, 10032, U.S.A
| | - Jérémy Chéret
- Dr. Philip Frost Department for Dermatology and Cutaneous Surgery, University of Miami, Miami, Florida, 33136, U.S.A
| | - Barbara Bedogni
- Dr. Philip Frost Department for Dermatology and Cutaneous Surgery, University of Miami, Miami, Florida, 33136, U.S.A
| | - Desmond J Tobin
- Charles Institute of Dermatology, University College Dublin, Dublin 4, Ireland
| | - Ralf Paus
- Dr. Philip Frost Department for Dermatology and Cutaneous Surgery, University of Miami, Miami, Florida, 33136, U.S.A.,Monasterium Laboratory, Skin & Hair Research Solutions GmbH, Münster, D-48149, Germany.,Centre for Dermatology Research, NIHR Manchester Biomedical Research Centre, University of Manchester, Manchester, M13 9PT, U.K
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Li H, Masieri FF, Schneider M, Kottek T, Hahnel S, Yamauchi K, Obradović D, Seon JK, Yun SJ, Ferrer RA, Franz S, Simon JC, Lethaus B, Savković V. Autologous, Non-Invasively Available Mesenchymal Stem Cells from the Outer Root Sheath of Hair Follicle Are Obtainable by Migration from Plucked Hair Follicles and Expandable in Scalable Amounts. Cells 2020; 9:E2069. [PMID: 32927740 PMCID: PMC7564264 DOI: 10.3390/cells9092069] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2020] [Revised: 09/04/2020] [Accepted: 09/06/2020] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Regenerative therapies based on autologous mesenchymal stem cells (MSC) as well as stem cells in general are still facing an unmet need for non-invasive sampling, availability, and scalability. The only known adult source of autologous MSCs permanently available with no pain, discomfort, or infection risk is the outer root sheath of the hair follicle (ORS). METHODS This study presents a non-invasively-based method for isolating and expanding MSCs from the ORS (MSCORS) by means of cell migration and expansion in air-liquid culture. RESULTS The method yielded 5 million cells of pure MSCORS cultured in 35 days, thereby superseding prior art methods of culturing MSCs from hair follicles. MSCORS features corresponded to the International Society for Cell Therapy characterization panel for MSCs: adherence to plastic, proliferation, colony forming, expression of MSC-markers, and adipo-, osteo-, and chondro-differentiation capacity. Additionally, MSCORS displayed facilitated random-oriented migration and high proliferation, pronounced marker expression, extended endothelial and smooth muscle differentiation capacity, as well as a paracrine immunomodulatory effect on monocytes. MSCORS matched or even exceeded control adipose-derived MSCs in most of the assessed qualities. CONCLUSIONS MSCORS qualify for a variety of autologous regenerative treatments of chronic disorders and prophylactic cryopreservation for purposes of acute treatments in personalized medicine.
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Affiliation(s)
- Hanluo Li
- Department of Cranial Maxillofacial Plastic Surgery, University Clinic Leipzig, 04103 Leipzig, Germany; (H.L.); (T.K.); (B.L.)
| | - Federica Francesca Masieri
- School of (EAST) Engineering, Arts, Science & Technology, University of Suffolk, Ipswich, Suffolk IP41QJ, UK;
| | - Marie Schneider
- Clinic for Hematology, Cell Therapy and Hemostaseology, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Tina Kottek
- Department of Cranial Maxillofacial Plastic Surgery, University Clinic Leipzig, 04103 Leipzig, Germany; (H.L.); (T.K.); (B.L.)
| | - Sebastian Hahnel
- Polyclinic for Dental Prosthetics and Material Sciences, University Hospital Leipzig, 04103 Leipzig, Germany;
| | - Kensuke Yamauchi
- Kensuke Yamauchi, Department of Oral & Maxillofacial Surgery, Tohoku University, Sendai 980-8577, Japan;
| | | | - Jong-Keun Seon
- Chonnam National University Hwasun Hospital, Hwasun-gun 58128, Korea; (J.-K.S.); (S.J.Y.)
| | - Sook Jung Yun
- Chonnam National University Hwasun Hospital, Hwasun-gun 58128, Korea; (J.-K.S.); (S.J.Y.)
| | - Rubén A. Ferrer
- Clinic for Dermatology, Venereology and Allergology, University Hospital Leipzig, 04103 Leipzig, Germany; (R.A.F.); (S.F.); (J.-C.S.)
| | - Sandra Franz
- Clinic for Dermatology, Venereology and Allergology, University Hospital Leipzig, 04103 Leipzig, Germany; (R.A.F.); (S.F.); (J.-C.S.)
| | - Jan-Christoph Simon
- Clinic for Dermatology, Venereology and Allergology, University Hospital Leipzig, 04103 Leipzig, Germany; (R.A.F.); (S.F.); (J.-C.S.)
| | - Bernd Lethaus
- Department of Cranial Maxillofacial Plastic Surgery, University Clinic Leipzig, 04103 Leipzig, Germany; (H.L.); (T.K.); (B.L.)
| | - Vuk Savković
- Department of Cranial Maxillofacial Plastic Surgery, University Clinic Leipzig, 04103 Leipzig, Germany; (H.L.); (T.K.); (B.L.)
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Obara K, Tohgi N, Shirai K, Mii S, Hamada Y, Arakawa N, Aki R, Singh SR, Hoffman RM, Amoh Y. Hair-Follicle-Associated Pluripotent (HAP) Stem Cells Encapsulated on Polyvinylidene Fluoride Membranes (PFM) Promote Functional Recovery from Spinal Cord Injury. Stem Cell Rev Rep 2020; 15:59-66. [PMID: 30341634 DOI: 10.1007/s12015-018-9856-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
Our previous studies showed that nestin-expressing hair follicle-associated-pluripotent (HAP) stem cells, which reside in the bulge area of the hair follicle, could restore injured nerve and spinal cord and differentiate into cardiac muscle cells. Here we transplanted mouse green fluorescent protein (GFP)-expressing HAP stem-cell colonies enclosed on polyvinylidene fluoride membranes (PFM) into the severed thoracic spinal cord of nude mice. After seven weeks of implantation, we found the differentiation of HAP stem cells into neurons and glial cells. Our results also showed that PFM-captured GFP-expressing HAP stem-cell colonies assisted complete reattachment of the thoracic spinal cord. Furthermore, our quantitative motor function analysis with the Basso Mouse Scale for Locomotion (BMS) score demonstrated a significant improvement in the implanted mice compared to non-implanted mice with a severed spinal cord. Our study also showed that it is easy to obtain HAP stem cells, they do not develop teratomas, and do not loose differentiation ability when cryopreserved. Collectively our results suggest that HAP stem cells could be a better source compared to induced pluripotent stem cells (iPS) or embryonic stem (ES) cells for regenerative medicine, specifically for spinal cord repair.
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Affiliation(s)
- Koya Obara
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Natsuko Tohgi
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Kyoumi Shirai
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Sumiyuki Mii
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Yuko Hamada
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Nobuko Arakawa
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Ryoichi Aki
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan
| | - Shree Ram Singh
- Basic Research Laboratory, National Cancer Institute, Frederick, MD, 21702, USA.
| | - Robert M Hoffman
- AntiCancer, Inc., 7917 Ostrow Street, San Diego, CA, 92111, USA. .,Department of Surgery, University of California, San Diego, CA, 92103, USA.
| | - Yasuyuki Amoh
- Department of Dermatology, Kitasato University School of Medicine, Minami Ward, Sagamihara, 252-0374, Japan.
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Dong D, Chen S, Feng C, Xiong H, Xu X. NB-UVB Induces Melanocytic Differentiation of Human Hair Follicle Neural Crest Stem Cells. Ann Dermatol 2020; 32:289-297. [PMID: 33911756 PMCID: PMC7992648 DOI: 10.5021/ad.2020.32.4.289] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/02/2019] [Revised: 03/15/2020] [Accepted: 03/16/2020] [Indexed: 11/08/2022] Open
Abstract
Background Phototherapy is an important method to treat vitiligo. However, it is unclear how phototherapy affects melanocyte precursors and skin neural crest stem cells. Objective To investigate the underlying mechanisms of narrow-band ultraviolet B (NB-UVB) induced melanocyte lineage differentiated from human scalp-derived neural crest stem cells (HS-NCSCs). Methods HS-NCSCs were expanded from scalp hair follicles. The c-Kit-/CD57- HS-NCSCs were isolated by cell sorting. Different doses of NB-UVB were used to irradiate these HS-NCSCs. Cell ultrastructure was examined by transmission electron microscope. Melanocyte marker expression was analyzed by Quantitative RT-PCR and Western blot. Cell proliferation and migration were also evaluated. Results The c-Kit-/CD57- HS-NCSCs expressed embryonic NCSC biomarkers. NB-UVB at a dose of 100 mJ of NB-UVB had little effect on the cell proliferation of differentiated melanocytes from c-Kit-/CD57- HS-NCSCs, while 700 mJ inhibited cell proliferation significantly. The dendritic processes of differentiated melanocytes increased after radiation. The tyrosinase and Melanocortin 1 receptor (Mc1R) expression of differentiated melanocytes increased after NB-UVB exposure. The effect of NB-UVB on tyrosinase expression was modulated by signaling inhibitors H89 and PD98059 as well as Mc1R level in the cells. The migration ability of differentiated melanocytes was enhanced under 100 mJ exposure. Conclusion These data demonstrate that NB-UVB facilitates melanocytic differentiation of the HS-NCSCs and enhances migration of these cells. Mc1R and cAMP pathway play a critical role in NB-UVB induced melanocytic differentiation.
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Affiliation(s)
- Dake Dong
- Department of Dermatology, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.,Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
| | - Shujun Chen
- Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China
| | - Cheng Feng
- Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.,Department of Dermatology, the Second Affiliated Hospital of Xi'anJiaoTong University, Xi'an, Shanxi, China
| | - Huizi Xiong
- Department of Dermatology, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China
| | - Xiaowei Xu
- Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
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Sweat gland regeneration: Current strategies and future opportunities. Biomaterials 2020; 255:120201. [PMID: 32592872 DOI: 10.1016/j.biomaterials.2020.120201] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2020] [Revised: 04/21/2020] [Accepted: 06/09/2020] [Indexed: 12/13/2022]
Abstract
For patients with extensive skin defects, loss of sweat glands (SwGs) greatly decreases their quality of life. Indeed, difficulties in thermoregulation, ion reabsorption, and maintaining fluid balance might render them susceptible to hyperthermia, heatstroke, or even death. Despite extensive studies on the stem cell biology of the skin in recent years, in-situ regeneration of SwGs with both structural and functional fidelity is still challenging because of the limited regenerative capacity and cell fate control of resident progenitors. To overcome these challenges, one must consider both the intrinsic factors relevant to genetic and epigenetic regulation and cues from the cellular microenvironment. Here, we describe recent progress in molecular biology, developmental pathways, and cellular evolution associated with SwGdevelopment and maturation. This is followed by a summary of the current strategies used for cell-fate modulation, transmembrane drug delivery, and scaffold design associated with SwGregeneration. Finally, we offer perspectives for creating more sophisticated systems to accelerate patients' innate healing capacity and developing engineered skin constructs to treat or replace damaged tissues structurally and functionally.
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Wang T, Lin H, Liu F, Zhang C. Olig2 positive cells derived from hair follicle neural crest stem cells in rats. J Chem Neuroanat 2020; 105:101770. [PMID: 32088378 DOI: 10.1016/j.jchemneu.2020.101770] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/28/2019] [Revised: 02/17/2020] [Accepted: 02/19/2020] [Indexed: 10/25/2022]
Abstract
Motor neuron disease (MND) is a kind of common clinical nervous system disease with typical characteristic of progressive motor neurons degeneration or death. Motor neuron derived from stem cells or motor neuron progenitor cells will be a good choice to be used for treatment of the disease. In this study, we used the combination of 5 small molecular including CHIR99021 (CHIR), SB431542 (SB), DMH1 (DMH), retinoic acid (RA) and Purmorphamine (Pur) to induce hair follicles neural crest stem cells (hfNCSCs) to motor neurons progenitors (MNPs). Valproic acid (VPA) was used to make MNPs proliferation. RA and Pur were used to try to induce MNPs toward motor neurons (MNs) and CpdE was tried for MNs maturation. Nestin, β-tubulin Ш (Tuj1), microtubule associated protein 2 (MAP2), Olig2, choline acetyltransferase (ChAT)and TUBB3 were examined at protein and mRNA levels by immunofluoresence cytochemistry, western blot and real time PCR at 6, 16 and 22 days. Our data showed cells changed into bipolar or multipolar shape forming the cell clusters like scattered rosettes. Nestin expression decreased significantly at 22 days. Compared to 6 days, percentage of Olig2 + MNPs was higher, (88.53 ± 6.67)%, and Olig2 expression at protein and gene level was lower at 22 days. Percentage of MAP2 positive cells increased to (90.62 ± 2.31) % and ChAT positive cells increased to (83.29 ± 6.62) % at 22 days. But no expression of ChAT was examined by western blot and real time PCR. It indicates that these 5 molecular can differentiate hfNCSCs into Olig2 positive cells with a unipotent differentiation toward motor neurons.
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Affiliation(s)
- Tao Wang
- Department of Anatomy, the Second Military Medical University/Naval Medical University, China
| | - Haiyan Lin
- Department of Anatomy, the Second Military Medical University/Naval Medical University, China.
| | - Fang Liu
- Department of Anatomy, the Second Military Medical University/Naval Medical University, China
| | - Chuansen Zhang
- Department of Anatomy, the Second Military Medical University/Naval Medical University, China
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Sumathy B, Nair PD. Keratinocytes-hair follicle bulge stem cells-fibroblasts co-cultures on a tri-layer skin equivalent derived from gelatin/PEG methacrylate nanofibers. JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION 2020; 31:869-894. [DOI: 10.1080/09205063.2020.1725861] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Affiliation(s)
- Babitha Sumathy
- Division of Tissue Engineering and Regeneration Technologies, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, Kerala, India
| | - Prabha D Nair
- Division of Tissue Engineering and Regeneration Technologies, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, Kerala, India
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Khalil S, Ariel Gru A, Saavedra AP. Cutaneous extramedullary haematopoiesis: Implications in human disease and treatment. Exp Dermatol 2019; 28:1201-1209. [DOI: 10.1111/exd.14013] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2018] [Revised: 06/26/2019] [Accepted: 07/11/2019] [Indexed: 12/17/2022]
Affiliation(s)
- Shadi Khalil
- Department of Dermatology University of Virginia School of Medicine Charlottesville Virginia
| | - Alejandro Ariel Gru
- Department of Pathology University of Virginia School of Medicine Charlottesville Virginia
| | - Arturo P. Saavedra
- Department of Dermatology University of Virginia School of Medicine Charlottesville Virginia
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