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Chu X, Zhou Z, Qian X, Shen H, Cheng H, Zhang J. Functional regeneration strategies of hair follicles: advances and challenges. Stem Cell Res Ther 2025; 16:77. [PMID: 39985119 PMCID: PMC11846195 DOI: 10.1186/s13287-025-04210-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: 11/23/2024] [Accepted: 01/29/2025] [Indexed: 02/24/2025] Open
Abstract
Hair follicles are essential appendages of human skin that function in protection, sensation, thermoregulation and social interactions. The multicellular components, particularly the dermal papilla, matrix and bulge housing stem cells, enable cyclic hair growth postnatally. However, miniaturization and loss of hair follicles can occur in the context of ageing, trauma and various alopecia-related diseases. Conventional treatments involve the redistribution of existing follicles, which may not be viable in patients lacking follicular resources. Recent progress in the comprehension of morphogenesis and the development of biomaterials has significantly advanced follicle reconstruction, incorporating organ germ assembling, stem cell induction and bioprinting techniques. Despite these advancements, fully restoring hair follicles remains challenging due to the complexities of replicating embryonic signals and sustaining growth cycles. Identifying suitable cell sources for clinical applications also presents a hurdle. Here, we retrospect the progress made in the field of hair follicle regeneration, aiming to offer an exhaustive analysis on the benefits and limitations of these methods, and to foster the development of innovative solutions.
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Affiliation(s)
- Xi Chu
- Department of Plastic and Cosmetic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, 261 Huansha Road, Hangzhou, 310000, Zhejiang, China
| | - Zhentao Zhou
- Department of Plastic and Cosmetic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, 261 Huansha Road, Hangzhou, 310000, Zhejiang, China
| | - Xifei Qian
- School of Medicine, Zhejiang Chinese Medical University, Hangzhou, 310000, Zhejiang, China
| | - Haiyan Shen
- Department of Plastic and Cosmetic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, 261 Huansha Road, Hangzhou, 310000, Zhejiang, China
| | - Hanxiao Cheng
- Department of Plastic and Cosmetic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, 261 Huansha Road, Hangzhou, 310000, Zhejiang, China
| | - Jufang Zhang
- Department of Plastic and Cosmetic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, 261 Huansha Road, Hangzhou, 310000, Zhejiang, China.
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2
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Brandes N, Hahn H, Uhmann A. CD4 expression controls epidermal stem cell balance. Sci Rep 2025; 15:4185. [PMID: 39905055 PMCID: PMC11794708 DOI: 10.1038/s41598-025-87915-7] [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: 09/23/2024] [Accepted: 01/22/2025] [Indexed: 02/06/2025] Open
Abstract
The balance of stem cell populations is essential for the maintenance, renewal, and repair of the mammalian epidermis. Here, we report that CD4, which is a typical marker of helper T cells, monocytes, macrophages, and dendritic cells, is also expressed on murine K5+ keratinocytes. Lineage tracing of CD4+ cells reveals that their epidermal progeny has self-renewal abilities and clonogenic potential. The progeny of CD4+ epidermal cells contributes to epidermal renewal and progressively colonizes the interfollicular epidermis and hair follicles with age, thereby developing to all epidermal lineages. Wound healing studies furthermore show that the progeny of CD4+ epidermal cells accumulates at wound sites. Finally, using CD4 knockout mice we demonstrate that CD4 expression is essential for maintaining fast-cycling epidermal stem cells during homeostasis and that CD4 loss mitigates the age-related decline in wound repair capacity. Collectively, our data support the conclusion that CD4 expression is required for long-term maintenance of the epidermal stem cell balance.
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Affiliation(s)
- Nadine Brandes
- Institute of Human Genetics, Tumor Genetics Group, Universitätsmedizin Göttingen, Heinrich-Düker-Weg 12, 37073, Göttingen, Germany
| | - Heidi Hahn
- Institute of Human Genetics, Tumor Genetics Group, Universitätsmedizin Göttingen, Heinrich-Düker-Weg 12, 37073, Göttingen, Germany
| | - Anja Uhmann
- Institute of Human Genetics, Tumor Genetics Group, Universitätsmedizin Göttingen, Heinrich-Düker-Weg 12, 37073, Göttingen, Germany.
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3
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Hamida OB, Kim MK, Sung YK, Kim MK, Kwack MH. Hair Regeneration Methods Using Cells Derived from Human Hair Follicles and Challenges to Overcome. Cells 2024; 14:7. [PMID: 39791708 PMCID: PMC11720663 DOI: 10.3390/cells14010007] [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: 11/15/2024] [Revised: 12/12/2024] [Accepted: 12/23/2024] [Indexed: 01/12/2025] Open
Abstract
The hair follicle is a complex of mesenchymal and epithelial cells acquiring different properties and characteristics responsible for fulfilling its inductive and regenerative role. The epidermal and dermal crosstalk induces morphogenesis and maintains hair follicle cycling properties. The hair follicle is enriched with pluripotent stem cells, where dermal papilla (DP) cells and dermal sheath (DS) cells constitute the dermal compartment and the epithelial stem cells existing in the bulge region exert their regenerative role by mediating the epithelial-mesenchymal interaction (EMI). Many studies have developed and focused on various methods to optimize the EMI through in vivo and in vitro approaches for hair regeneration. The culturing of human hair mesenchymal cells resulted in the loss of trichogenicity and inductive properties of DP cells, limiting their potential application in de novo hair follicle generation in vivo. Epithelial stem cells derived from human hair follicles are challenging to isolate and culture, making it difficult to obtain enough cells for hair regeneration purposes. Mesenchymal stem cells and epithelial stem cells derived from human hair follicles lose their ability to form hair follicles during culture, limiting the study of hair follicle formation in vivo. Therefore, many attempts and methods have been developed to overcome these limitations. Here, we review the possible and necessary cell methods and techniques used for human hair follicle regeneration and the restoration of hair follicle cell inductivity in culture.
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Affiliation(s)
- Ons Ben Hamida
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea; (O.B.H.); (M.K.K.); (Y.K.S.); (M.K.K.)
| | - Moon Kyu Kim
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea; (O.B.H.); (M.K.K.); (Y.K.S.); (M.K.K.)
- Hair Transplantation Center, Kyungpook National University Hospital, Daegu 41944, Republic of Korea
| | - Young Kwan Sung
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea; (O.B.H.); (M.K.K.); (Y.K.S.); (M.K.K.)
| | - Min Kyu Kim
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea; (O.B.H.); (M.K.K.); (Y.K.S.); (M.K.K.)
| | - Mi Hee Kwack
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea; (O.B.H.); (M.K.K.); (Y.K.S.); (M.K.K.)
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4
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Moriwaki Y, Shiraishi M, Shen Q, Du Z, Okazaki M, Kurita M. Experimental method for creating skin with acquired appendage dysfunction. J Dermatol 2024. [PMID: 39676456 DOI: 10.1111/1346-8138.17579] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2024] [Revised: 11/16/2024] [Accepted: 11/28/2024] [Indexed: 12/17/2024]
Abstract
Mammalian skin appendages, such as hair follicles and sweat glands, are essential for both esthetic and functional purposes. Conditions such as burns and ulcers can lead to dysfunction or loss of skin appendages and result in hair loss and dry skin, posing challenges in their regeneration. Existing animal models are insufficient for studying acquired dysfunction of skin appendages without underlying genetic causes. This study aimed to develop more clinically relevant mouse models by evaluating two approaches: keratinocyte transplantation and grafting of skin at varying thicknesses. green fluorescent protein (GFP)-expressing keratinocytes were transplanted into ulcers on nude mice, leading to re-epithelialization with minimal skin appendages at 4 weeks after transplantation. However, the re-epithelialized area was largely derived from recipient cells, with the grafted cells contributing to only 1.31% of the area. In the skin-grafting model, donor skin from GFP transgenic mice was grafted onto nude mice at three thicknesses: full thickness, 10/1000 inch, and 5/1000 inch. The grafted area of the 5/1000-inch grafts remained stable at 89.5% of its original size 5 weeks after transplantation, ensuring a sufficiently large skin area. The 5/1000-inch grafts resulted in a significant reduction in skin appendages, with a mean of only 3.73 hair follicles per 5 mm, compared with 69.7 in the control group. The 5/1000-inch skin grafting in orthotopic autologous transplantation also showed the achievement of skin surfaces with a minimal number of skin appendages. Therefore, a mouse model with skin grafting demonstrated stability in producing large areas of skin with minimal appendages. In conclusion, these two models with acquired skin appendage dysfunction and no underlying genetic causes provide valuable tools for researching skin appendage regeneration, offering insights into potential therapeutic strategies for conditions involving skin appendage loss.
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Affiliation(s)
- Yuta Moriwaki
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
| | - Makoto Shiraishi
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
| | - Qi Shen
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
| | - Zening Du
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
| | - Mutsumi Okazaki
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
| | - Masakazu Kurita
- Department of Plastic and Reconstructive Surgery, The University of Tokyo Hospital, Tokyo, Japan
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5
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Tang T, Zhang P, Zhang Q, Man X, Xu Y. Fabrication of heterocellular spheroids with controllable core-shell structure using inertial focusing effect for scaffold-free 3D cell culture models. Biofabrication 2024; 16:045013. [PMID: 39019062 DOI: 10.1088/1758-5090/ad647e] [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: 03/31/2024] [Accepted: 07/17/2024] [Indexed: 07/19/2024]
Abstract
Three-dimensional (3D) cell culture models capable of emulating the biological functions of natural tissues are pivotal in tissue engineering and regenerative medicine. Despite progress, the fabrication ofin vitroheterocellular models that mimic the intricate structures of natural tissues remains a significant challenge. In this study, we introduce a novel, scaffold-free approach leveraging the inertial focusing effect in rotating hanging droplets for the reliable production of heterocellular spheroids with controllable core-shell structures. Our method offers precise control over the core-shell spheroid's size and geometry by adjusting the cell suspension density and droplet morphology. We successfully applied this technique to create hair follicle organoids, integrating dermal papilla cells within the core and epidermal cells in the shell, thereby achieving markedly enhanced hair inducibility compared to mixed-structure models. Furthermore, we have developed melanoma tumor spheroids that accurately mimic the dynamic interactions between tumor and stromal cells, showing increased invasion capabilities and altered expressions of cellular adhesion molecules and proteolytic enzymes. These findings underscore the critical role of cellular spatial organization in replicating tissue functionalityin vitro. Our method represents a significant advancement towards generating heterocellular spheroids with well-defined architectures, offering broad implications for biological research and applications in tissue engineering.
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Affiliation(s)
- Tan Tang
- School of Mechanical Engineering and Automation, Beihang University, Beijing, People's Republic of China
| | - Pengfei Zhang
- School of Mechanical Engineering and Automation, Beihang University, Beijing, People's Republic of China
| | - Qiuting Zhang
- School of Mechanical Engineering and Automation, Beihang University, Beijing, People's Republic of China
| | - Xingkun Man
- School of Physics, Beihang University, Beijing, People's Republic of China
| | - Ye Xu
- School of Mechanical Engineering and Automation, Beihang University, Beijing, People's Republic of China
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6
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Ramos R, Swedlund B, Ganesan AK, Morsut L, Maini PK, Monuki ES, Lander AD, Chuong CM, Plikus MV. Parsing patterns: Emerging roles of tissue self-organization in health and disease. Cell 2024; 187:3165-3186. [PMID: 38906093 PMCID: PMC11299420 DOI: 10.1016/j.cell.2024.05.016] [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: 09/12/2023] [Revised: 02/22/2024] [Accepted: 05/08/2024] [Indexed: 06/23/2024]
Abstract
Patterned morphologies, such as segments, spirals, stripes, and spots, frequently emerge during embryogenesis through self-organized coordination between cells. Yet, complex patterns also emerge in adults, suggesting that the capacity for spontaneous self-organization is a ubiquitous property of biological tissues. We review current knowledge on the principles and mechanisms of self-organized patterning in embryonic tissues and explore how these principles and mechanisms apply to adult tissues that exhibit features of patterning. We discuss how and why spontaneous pattern generation is integral to homeostasis and healing of tissues, illustrating it with examples from regenerative biology. We examine how aberrant self-organization underlies diverse pathological states, including inflammatory skin disorders and tumors. Lastly, we posit that based on such blueprints, targeted engineering of pattern-driving molecular circuits can be leveraged for synthetic biology and the generation of organoids with intricate patterns.
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Affiliation(s)
- Raul Ramos
- Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, USA; NSF-Simons Center for Multiscale Cell Fate Research, University of California, Irvine, Irvine, CA, USA
| | - Benjamin Swedlund
- Eli and Edythe Broad CIRM Center, Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA
| | - Anand K Ganesan
- Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA; Department of Dermatology, University of California, Irvine, Irvine, CA, USA
| | - Leonardo Morsut
- Eli and Edythe Broad CIRM Center, Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA; Alfred E. Mann Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA
| | - Philip K Maini
- Mathematical Institute, University of Oxford, Oxford, UK
| | - Edwin S Monuki
- Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, USA; Department of Pathology and Laboratory Medicine, University of California, Irvine, Irvine, CA, USA
| | - Arthur D Lander
- Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA; Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA.
| | - Cheng-Ming Chuong
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
| | - Maksim V Plikus
- Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, USA; NSF-Simons Center for Multiscale Cell Fate Research, University of California, Irvine, Irvine, CA, USA; Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA.
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7
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Sugiyama E, Nanmo A, Nie X, Chang SY, Hashimoto M, Suzuki A, Kageyama T, Fukuda J. Large-Scale Preparation of Hair Follicle Germs Using a Microfluidic Device. ACS Biomater Sci Eng 2024; 10:998-1005. [PMID: 38193447 PMCID: PMC10865290 DOI: 10.1021/acsbiomaterials.3c01346] [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: 09/17/2023] [Revised: 12/13/2023] [Accepted: 12/13/2023] [Indexed: 01/10/2024]
Abstract
Hair follicle morphogenesis during embryonic development is driven by the formation of hair follicle germs (HFGs) via interactions between epithelial and mesenchymal cells. Bioengineered HFGs are potential tissue grafts for hair regenerative medicine because they can replicate interactions and hair follicle morphogenesis after transplantation. However, a mass preparation approach for HFGs is necessary for clinical applications, given that thousands of de novo hair follicles are required to improve the appearance of a single patient with alopecia. In this study, we developed a microfluidics-based approach for the large-scale preparation of HFGs. A simple flow-focusing microfluidic device allowed collagen solutions containing epithelial and mesenchymal cells to flow and generate collagen microbeads with distinct Janus structures. During the 3 days of culture, the collagen beads contracted owing to cellular traction forces, resulting in collagen- and cell-dense HFGs. The transplantation of HFGs into nude mice resulted in highly efficient de novo hair follicle regeneration. This method provides a scalable and robust tissue graft preparation approach for hair regeneration.
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Affiliation(s)
- Ellen Sugiyama
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Ayaka Nanmo
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Xiaolei Nie
- Pillar
of Engineering Product Development, Singapore
University of Technology and Design, 8 Somapah Road, Singapore 487372, Singapore
- Digital
Manufacturing and Design (DManD) Centre, Singapore University of Technology and Design, 8 Somapah Rd, Singapore 487372, Singapore
| | - Shu-Yung Chang
- Pillar
of Engineering Product Development, Singapore
University of Technology and Design, 8 Somapah Road, Singapore 487372, Singapore
- Digital
Manufacturing and Design (DManD) Centre, Singapore University of Technology and Design, 8 Somapah Rd, Singapore 487372, Singapore
| | - Michinao Hashimoto
- Pillar
of Engineering Product Development, Singapore
University of Technology and Design, 8 Somapah Road, Singapore 487372, Singapore
- Digital
Manufacturing and Design (DManD) Centre, Singapore University of Technology and Design, 8 Somapah Rd, Singapore 487372, Singapore
| | - Atsushi Suzuki
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Institute
of Advanced Sciences, Yokohama National
University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Tatsuto Kageyama
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Institute
of Advanced Sciences, Yokohama National
University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Kanagawa
Institute of Industrial Science and Technology, 3-2-1 Sakado Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan
| | - Junji Fukuda
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Institute
of Advanced Sciences, Yokohama National
University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Kanagawa
Institute of Industrial Science and Technology, 3-2-1 Sakado Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan
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8
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Zhu N, Yan J, Gu W, Yang Q, Lin E, Lu S, Cai B, Xia B, Liu X, Lin C. Dermal papilla cell-secreted biglycan regulates hair follicle phase transit and regeneration by activating Wnt/β-catenin. Exp Dermatol 2024; 33:e14969. [PMID: 37967213 DOI: 10.1111/exd.14969] [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: 03/28/2023] [Revised: 09/06/2023] [Accepted: 10/19/2023] [Indexed: 11/17/2023]
Abstract
Alopecia is a prevalent problem of cutaneous appendages and lacks effective therapy. Recently, researchers have been focusing on mesenchymal components of the hair follicle, i.e. dermal papilla cells, and we previously identified biglycan secreted by dermal papilla cells as the key factor responsible for hair follicle-inducing ability. In this research, we hypothesized biglycan played an important role in hair follicle cycle and regeneration through regulating the Wnt signalling pathway. To characterize the hair follicle cycle and the expression pattern of biglycan, we observed hair follicle morphology in C57BL/6 mice on Days 0, 3, 5, 12 and 18 post-depilation and found that biglycan is highly expressed at both mRNA and protein levels throughout anagen in HFs. To explore the role of biglycan during the phase transit process and regeneration, local injections were administered in C57BL/6 and nude mice. Results showed that local injection of biglycan in anagen HFs delayed catagen progression and involve activating the Wnt/β-catenin signalling pathway. Furthermore, local injection of biglycan induced HF regeneration and up-regulated expression of key Wnt factors in nude mice. In addition, cell analyses exhibited biglycan knockdown inactivated the Wnt signalling pathway in early-passage dermal papilla cell, whereas biglycan overexpression or incubation activated the Wnt signalling pathway in late-passage dermal papilla cells. These results indicate that biglycan plays a critical role in regulating HF cycle transit and regeneration in a paracrine and autocrine fashion by activating the Wnt/β-catenin signalling pathway and could be a potential treatment target for hair loss diseases.
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Affiliation(s)
- Ningxia Zhu
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Junping Yan
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Weifan Gu
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Qilin Yang
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - En Lin
- Department of Histology and Embryology, Shantou University Medical College, Shantou, People's Republic of China
| | - Siyue Lu
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Bozhi Cai
- Tissue Engineering Laboratory, First Affiliated Hospital, Shantou University Medical College, Shantou, People's Republic of China
| | - Bin Xia
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Xin Liu
- Department of Pathology and Physiopathology, Guilin Medical University, Guilin, People's Republic of China
| | - Changmin Lin
- Department of Histology and Embryology, Shantou University Medical College, Shantou, People's Republic of China
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9
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Lei M, Jiang J, Wang M, Wu W, Zhang J, Liu W, Zhou W, Lai YC, Jiang TX, Widelitz RB, Harn HIC, Yang L, Chuong CM. Epidermal-dermal coupled spheroids are important for tissue pattern regeneration in reconstituted skin explant cultures. NPJ Regen Med 2023; 8:65. [PMID: 37996466 PMCID: PMC10667216 DOI: 10.1038/s41536-023-00340-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/01/2022] [Accepted: 11/09/2023] [Indexed: 11/25/2023] Open
Abstract
Tissue patterning is critical for the development and regeneration of organs. To advance the use of engineered reconstituted skin organs, we study cardinal features important for tissue patterning and hair regeneration. We find they spontaneously form spheroid configurations, with polarized epidermal cells coupled with dermal cells through a newly formed basement membrane. Functionally, the spheroid becomes competent morphogenetic units (CMU) that promote regeneration of tissue patterns. The emergence of new cell types and molecular interactions during CMU formation was analyzed using scRNA-sequencing. Surprisingly, in newborn skin explants, IFNr signaling can induce apical-basal polarity in epidermal cell aggregates. Dermal-Tgfb induces basement membrane formation. Meanwhile, VEGF signaling mediates dermal cell attachment to the epidermal cyst shell, thus forming a CMU. Adult mouse and human fetal scalp cells fail to form a CMU but can be restored by adding IFNr or VEGF to achieve hair regeneration. We find different multi-cellular configurations and molecular pathways are used to achieve morphogenetic competence in developing skin, wound-induced hair neogenesis, and reconstituted explant cultures. Thus, multiple paths can be used to achieve tissue patterning. These insights encourage more studies of "in vitro morphogenesis" which may provide novel strategies to enhance regeneration.
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Affiliation(s)
- Mingxing Lei
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China.
- Integrative Stem Cell Center, China Medical University Hospital, China Medical University, Taichung, 40402, Taiwan.
| | - Jingwei Jiang
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Mengyue Wang
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Wang Wu
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Jinwei Zhang
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Wanqian Liu
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Wei Zhou
- Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, 400030, China
| | - Yung-Chih Lai
- Integrative Stem Cell Center, China Medical University Hospital, China Medical University, Taichung, 40402, Taiwan
| | - Ting-Xin Jiang
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA
| | - Randall B Widelitz
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA
| | - Hans I-Chen Harn
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA
| | - Li Yang
- Key Laboratory of Biorheological Science and Technology of Ministry of Education & 111 Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China
| | - Cheng-Ming Chuong
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA.
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10
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Sumathy B, Velayudhan S. Fabrication and evaluation of a bi-layered gelatin based scaffold with arrayed micro-pits for full-thickness skin construct. Int J Biol Macromol 2023; 251:126360. [PMID: 37591428 DOI: 10.1016/j.ijbiomac.2023.126360] [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: 04/29/2023] [Revised: 05/17/2023] [Accepted: 08/14/2023] [Indexed: 08/19/2023]
Abstract
There is an unmet need for a reliable and reproducible method for incorporating hair follicle derived stem cells in tissue engineered skin models to reconstitute hair follicles. This study discloses a novel method for introducing hair follicle derived stem cells in microneedle embossed micro-pits of a bilayer skin equivalent fabricated from a gelatin based scaffold. The microneedles are hard and strong enough to penetrate the upper layer of the bilayer gelatin based scaffold that corresponds to the epidermis and permeates down to lower layer that corresponds to dermal layer. This strategic location will mimic the natural niche of hair follicle stem cells for picking up signals from both the epidermis and dermis. Hair follicle stem cells are trapped in to these micro-pits by vacuum assisted cell seeding. The bilayer system consists of two distinct electrospun layers in a single processing step, representing outer epidermal layer and inner dermal layer with hair follicle stem cells in embedded pits, resulting in the formation of a closed representation of a complete skin.
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Affiliation(s)
- Babitha Sumathy
- Department of Tissue Engineering and Regeneration Technologies, Department of Applied Biology, Biomedical Technology wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum 695 012, India.
| | - Shiny Velayudhan
- Division of Dental Products, Department of Biomaterials Science and Technology, Biomedical Technology wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum 695 012, India.
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11
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Liu WX, Li CX, Xie XX, Ge W, Qiao T, Sun XF, Shen W, Cheng SF. Transcriptomic landscape reveals germline potential of porcine skin-derived multipotent dermal fibroblast progenitors. Cell Mol Life Sci 2023; 80:224. [PMID: 37480481 PMCID: PMC11072884 DOI: 10.1007/s00018-023-04869-7] [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: 03/16/2023] [Revised: 06/15/2023] [Accepted: 07/10/2023] [Indexed: 07/24/2023]
Abstract
According to estimations, approximately about 15% of couples worldwide suffer from infertility, in which individuals with azoospermia or oocyte abnormalities cannot be treated with assisted reproductive technology. The skin-derived stem cells (SDSCs) differentiation into primordial germ cell-like cells (PGCLCs) is one of the major breakthroughs in the field of stem cells intervention for infertility treatment in recent years. However, the cellular origin of SDSCs and their dynamic changes in transcription profile during differentiation into PGCLCs in vitro remain largely undissected. Here, the results of single-cell RNA sequencing indicated that porcine SDSCs are mainly derived from multipotent dermal fibroblast progenitors (MDFPs), which are regulated by growth factors (EGF/bFGF). Importantly, porcine SDSCs exhibit pluripotency for differentiating into three germ layers and can effectively differentiate into PGCLCs through complex transcriptional regulation involving histone modification. Moreover, this study also highlights that porcine SDSC-derived PGCLCs specification exhibit conservation with the human primordial germ cells lineage and that its proliferation is mediated by the MAPK signaling pathway. Our findings provide substantial novel insights into the field of regenerative medicine in which stem cells differentiate into germ cells in vitro, as well as potential therapeutic effects in individuals with azoospermia and/or defective oocytes.
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Affiliation(s)
- Wen-Xiang Liu
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China
- State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Sciences, Inner Mongolia University, Hohhot, 010021, China
| | - Chun-Xiao Li
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China
| | - Xin-Xiang Xie
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China
| | - Wei Ge
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China
| | - Tian Qiao
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China
| | - Xiao-Feng Sun
- Anqiu Women and Children's Hospital, Weifang, 262100, China
| | - Wei Shen
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China.
| | - Shun-Feng Cheng
- College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China.
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12
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Abstract
Pathological hair loss (also known as alopecia) and shortage of hair follicle (HF) donors have posed an urgent requirement for HF regeneration. With the revelation of mechanisms in tissue engineering, the proliferation of HFs in vitro has achieved more promising trust for the treatments of alopecia and other skin impairments. Theoretically, HF organoids have great potential to develop into native HFs and attachments such as sweat glands after transplantation. However, since the rich extracellular matrix (ECM) deficiency, the induction characteristics of skin-derived cells gradually fade away along with their trichogenic capacity after continuous cell passaging in vitro. Therefore, ECM-mimicking support is an essential prelude before HF transplantation is implemented. This review summarizes the status of providing various epidermal and dermal cells with a three-dimensional (3D) scaffold to support the cell homeostasis and better mimic in vivo environments for the sake of HF regeneration. HF-relevant cells including dermal papilla cells (DPCs), hair follicle stem cells (HFSCs), and mesenchymal stem cells (MSCs) are able to be induced to form HF organoids in the vitro culture system. The niche microenvironment simulated by different forms of biomaterial scaffold can offer the cells a network of ordered growth environment to alleviate inductivity loss and promote the expression of functional proteins. The scaffolds often play the role of ECM substrates and bring about epithelial-mesenchymal interaction (EMI) through coculture to ensure the functional preservation of HF cells during in vitro passage. Functional HF organoids can be formed either before or after transplantation into the dermis layer. Here, we review and emphasize the importance of 3D culture in HF regeneration in vitro. Finally, the latest progress in treatment trials and critical analysis of the properties and benefits of different emerging biomaterials for HF regeneration along with the main challenges and prospects of HF regenerative approaches are discussed.
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Affiliation(s)
- Wei Zheng
- College of Food Science & Technology, Shanghai Ocean University, Shanghai 201306, P.R. China
| | - Chang-Hua Xu
- College of Food Science & Technology, Shanghai Ocean University, Shanghai 201306, P.R. China
- Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China
- Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China
- National R&D Branch Center for Freshwater Aquatic Products Processing Technology (Shanghai), Shanghai 201306, China
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13
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Suzuki K, Yamaga K, Tokumasu R, Katsuno T, Tanaka H, Chiba S, Yagi T, Katayama I, Tamura A, Murota H, Tsukita S. Double mutation of claudin‐1 and claudin‐3 causes alopecia in infant mice. Ann N Y Acad Sci 2023; 1523:51-61. [PMID: 37002535 DOI: 10.1111/nyas.14980] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/03/2023]
Abstract
Hair follicles (HFs) undergo cyclic phases of growth, regression, and rest in association with hair shafts to maintain the hair coat. Nonsense mutations in the tight junction protein claudin (CLDN)-1 cause hair loss in humans. Therefore, we evaluated the roles of CLDNs in hair retention. Among the 27 CLDN family members, CLDN1, CLDN3, CLDN4, CLDN6, and CLDN7 were expressed in the inner bulge layer, isthmus, and sebaceous gland of murine HFs. Hair phenotypes were observed in Cldn1 weaker knockdown and Cldn3-knockout (Cldn1Δ/Δ Cldn3-/- ) mice. Although hair growth was normal, Cldn1Δ/Δ Cldn3-/- mice showed striking hair loss in the first telogen. Simultaneous deficiencies in CLDN1 and CLDN3 caused abnormalities in telogen HFs, such as an aberrantly layered architecture of epithelial cell sheets in bulges with multiple cell layers, mislocalization of bulges adjacent to sebaceous glands, and dilated hair canals. Along with the telogen HF abnormalities, which shortened the hair retention period, there was an enhanced proliferation of the epithelium surrounding HFs in Cldn1Δ/Δ Cldn3-/- mice, causing accelerated hair regrowth in adults. Our findings suggested that CLDN1 and CLDN3 may regulate hair retention in infant mice by maintaining the appropriate layered architecture of HFs, a deficiency of which can lead to alopecia.
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Affiliation(s)
- Koya Suzuki
- Advanced Comprehensive Research Organization Teikyo University Tokyo Japan
- Department of Clinical Laboratory of Medicine, Graduate School of Medicine Juntendo University Tokyo Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
| | - Kosuke Yamaga
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
- Department of Dermatology, Graduate School of Medicine Osaka University Osaka Japan
| | - Reitaro Tokumasu
- Advanced Comprehensive Research Organization Teikyo University Tokyo Japan
| | - Tatsuya Katsuno
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
- Center for Anatomical, Pathological and Forensic Medical Researches Kyoto University Graduate School of Medicine Kyoto Japan
- KOKORO‐Biology Group, Graduate School of Frontier Biosciences Osaka University Osaka Japan
| | - Hiroo Tanaka
- Advanced Comprehensive Research Organization Teikyo University Tokyo Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
- Department of Pharmacology Teikyo University School of Medicine Tokyo Japan
| | - Shuhei Chiba
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
- Laboratory of Molecular and Cellular Biology, Department of Biomolecular Sciences, Graduate School of Life Sciences Tohoku University Sendai Japan
| | - Takeshi Yagi
- KOKORO‐Biology Group, Graduate School of Frontier Biosciences Osaka University Osaka Japan
| | - Ichiro Katayama
- Department of Dermatology, Graduate School of Medicine Osaka University Osaka Japan
- Department of Pigmentation Research and Therapeutics, Graduate School of Medicine Osaka Metropolitan University Osaka Japan
| | - Atsushi Tamura
- Advanced Comprehensive Research Organization Teikyo University Tokyo Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
- Department of Pharmacology Teikyo University School of Medicine Tokyo Japan
| | - Hiroyuki Murota
- Department of Dermatology, Graduate School of Medicine Osaka University Osaka Japan
- Department of Dermatology Nagasaki University Graduate School of Biomedical Sciences Nagasaki Japan
| | - Sachiko Tsukita
- Advanced Comprehensive Research Organization Teikyo University Tokyo Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences Osaka University Osaka Japan
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14
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Hirano S, Kageyama T, Yamanouchi M, Yan L, Suzuki K, Ebisawa K, Kasai K, Fukuda J. Expansion Culture of Hair Follicle Stem Cells through Uniform Aggregation in Microwell Array Devices. ACS Biomater Sci Eng 2023; 9:1510-1519. [PMID: 36781164 PMCID: PMC10015430 DOI: 10.1021/acsbiomaterials.2c01141] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/15/2023]
Abstract
Hair regeneration using hair follicle stem cells (HFSCs) and dermal papilla cells is a promising approach for the treatment of alopecia. One of the challenges faced in this approach is the quantitative expansion of HFSCs while maintaining their hair induction capacity. In this study, HFSC expansion was achieved through the formation of uniform-diameter cell aggregates that were subsequently encapsulated in Matrigel. We designed a microwell array device, wherein mouse HFSCs were seeded, allowed to form loosely packed aggregates for an hour, and then embedded in Matrigel. Quantitative analysis revealed a 20-fold increase in HFSC number in 2 weeks through this culture device. Gene expression of trichogenic stem cell markers in the device-grown cells showed a significant increase compared with that of typical flat substrate Matrigel suspension culture cells. These microwell array-cultured HFSCs mixed with freshly isolated embryonic mesenchymal cells indicated vigorous hair regeneration on the skin of nude mice. Furthermore, we examined the feasibility of this approach for the expansion of human HFSCs from androgenetic alopecia patients and found that the ratio of CD200+ cells was improved significantly in comparison with that of cells cultured in a typical culture dish or in a Matrigel suspension culture on a flat substrate. Therefore, the novel approach proposed in this study may be useful for HFSC expansion in hair regenerative medicine.
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Affiliation(s)
- Sugi Hirano
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Tatsuto Kageyama
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Kanagawa
Institute of Industrial Science and Technology, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan
| | - Maki Yamanouchi
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Lei Yan
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Kanagawa
Institute of Industrial Science and Technology, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan
| | - Kohei Suzuki
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Nissan
Chemical Corporation, 2-5-1 Nihonbashi, Chuo-ku, Tokyo 103-6119, Japan
| | - Katsumi Ebisawa
- Department
of Plastic and Reconstructive Surgery, Nagoya
University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 464-8560, Japan
| | - Keiichiro Kasai
- Shonan
Beauty Clinic, 2-2-13
Yoyogi, Shibuya-ku, Tokyo 151-0053, Japan
| | - Junji Fukuda
- Faculty
of Engineering, Yokohama National University, 79-5 Tokiwadai,
Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
- Kanagawa
Institute of Industrial Science and Technology, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan
- . Tel: +81-45-339-4008. Fax: +81-45-339-4008
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15
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Huang J, Fu D, Wu X, Li Y, Zheng B, Liu Z, Zhou Y, Gan Y, Miao Y, Hu Z. One-step generation of core-shell biomimetic microspheres encapsulating double-layer cells using microfluidics for hair regeneration. Biofabrication 2023; 15. [PMID: 36608335 DOI: 10.1088/1758-5090/acb107] [Citation(s) in RCA: 10] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2022] [Accepted: 01/06/2023] [Indexed: 01/07/2023]
Abstract
Tissue engineering of hair follicles (HFs) has enormous potential in the treatment of hair loss. HF morphogenesis is triggered by reciprocal interactions between HF germ epithelial and mesenchymal layers. Here, a microfluidic-assisted technology is developed for the preparation of double aqueous microdroplets that entrap double-layer cells and growth factors to ultimately be used for hair regeneration. Mouse mesenchymal cells (MSCs) and epidermal cells (EPCs) are encapsulated in gelatin methacrylate (GelMA) cores and photo-curable catechol-grafted hyaluronic acid (HAD) shells to fabricate GelMA-MSC/HAD-EPC (G/HAD) microspheres. The findings show that the G/HAD microspheres exhibit ultrafast gelation, aqueous phase separation, superior biocompatibility, and favorable wet adhesion properties. G/HAD microspheres can also support cell proliferation and sustain growth factor release. These composite cell microspheres are capable of efficient HF generation upon transplantation into the dorsal dermis of nude mice. This finding facilitates the large-scale preparation of approximately 80 double-layer cell spheres per min. This simple double-layer cell sphere preparation approach is a promising strategy for improving current hair-regenerative medicine techniques and can potentially be applied along with other organoid techniques for extended applications.
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Affiliation(s)
- Junfei Huang
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Danlan Fu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Xiaoqi Wu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Yue Li
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - BoWen Zheng
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Zhen Liu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Yi Zhou
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Yuyang Gan
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Yong Miao
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
| | - Zhiqi Hu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, People's Republic of China
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16
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Aldea D, Kokalari B, Atsuta Y, Dingwall HL, Zheng Y, Nace A, Cotsarelis G, Kamberov YG. Differential modularity of the mammalian Engrailed 1 enhancer network directs sweat gland development. PLoS Genet 2023; 19:e1010614. [PMID: 36745673 PMCID: PMC9934363 DOI: 10.1371/journal.pgen.1010614] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/21/2022] [Revised: 02/16/2023] [Accepted: 01/13/2023] [Indexed: 02/07/2023] Open
Abstract
Enhancers are context-specific regulators of expression that drive biological complexity and variation through the redeployment of conserved genes. An example of this is the enhancer-mediated control of Engrailed 1 (EN1), a pleiotropic gene whose expression is required for the formation of mammalian eccrine sweat glands. We previously identified the En1 candidate enhancer (ECE) 18 cis-regulatory element that has been highly and repeatedly derived on the human lineage to potentiate ectodermal EN1 and induce our species' uniquely high eccrine gland density. Intriguingly, ECE18 quantitative activity is negligible outside of primates and ECE18 is not required for En1 regulation and eccrine gland formation in mice, raising the possibility that distinct enhancers have evolved to modulate the same trait. Here we report the identification of the ECE20 enhancer and show it has conserved functionality in mouse and human developing skin ectoderm. Unlike ECE18, knock-out of ECE20 in mice reduces ectodermal En1 and eccrine gland number. Notably, we find ECE20, but not ECE18, is also required for En1 expression in the embryonic mouse brain, demonstrating that ECE20 is a pleiotropic En1 enhancer. Finally, that ECE18 deletion does not potentiate the eccrine phenotype of ECE20 knock-out mice supports the secondary incorporation of ECE18 into the regulation of this trait in primates. Our findings reveal that the mammalian En1 regulatory machinery diversified to incorporate both shared and lineage-restricted enhancers to regulate the same phenotype, and also have implications for understanding the forces that shape the robustness and evolvability of developmental traits.
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Affiliation(s)
- Daniel Aldea
- Department of Genetics, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - Blerina Kokalari
- Department of Genetics, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - Yuji Atsuta
- Genetics Department, Harvard Medical School, Boston, Massachusetts, United States of America
| | - Heather L. Dingwall
- Department of Genetics, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - Ying Zheng
- Department of Dermatology, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - Arben Nace
- Department of Dermatology, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - George Cotsarelis
- Department of Dermatology, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
| | - Yana G. Kamberov
- Department of Genetics, Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America
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17
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Liu Z, Huang J, Kang D, Zhou Y, Du L, Qu Q, Wang J, Wen L, Fu D, Hu Z, Miao Y. Microenvironmental Reprogramming of Human Dermal Papilla Cells for Hair Follicle Tissue Engineering. Acta Biomater 2022:S1742-7061(22)00730-9. [DOI: 10.1016/j.actbio.2022.11.004] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/18/2022] [Revised: 10/16/2022] [Accepted: 11/02/2022] [Indexed: 11/08/2022]
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18
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Beheshtizadeh N, Gharibshahian M, Pazhouhnia Z, Rostami M, Zangi AR, Maleki R, Azar HK, Zalouli V, Rajavand H, Farzin A, Lotfibakhshaiesh N, Sefat F, Azami M, Webster TJ, Rezaei N. Commercialization and regulation of regenerative medicine products: Promises, advances and challenges. Biomed Pharmacother 2022; 153:113431. [DOI: 10.1016/j.biopha.2022.113431] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/07/2022] [Revised: 07/04/2022] [Accepted: 07/14/2022] [Indexed: 11/02/2022] Open
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19
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Wang X, Liu Y, He J, Wang J, Chen X, Yang R. Regulation of signaling pathways in hair follicle stem cells. BURNS & TRAUMA 2022; 10:tkac022. [PMID: 35795256 PMCID: PMC9250793 DOI: 10.1093/burnst/tkac022] [Citation(s) in RCA: 40] [Impact Index Per Article: 13.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/18/2021] [Revised: 02/07/2022] [Indexed: 11/21/2022]
Abstract
Hair follicle stem cells (HFSCs) reside in the bulge region of the outer root sheath of the hair follicle. They are considered slow-cycling cells that are endowed with multilineage differentiation potential and superior proliferative capacity. The normal morphology and periodic growth of HFSCs play a significant role in normal skin functions, wound repair and skin regeneration. The HFSCs involved in these pathophysiological processes are regulated by a series of cell signal transduction pathways, such as lymphoid enhancer factor/T-cell factor, Wnt/β-catenin, transforming growth factor-β/bone morphogenetic protein, Notch and Hedgehog. The mechanisms of the interactions among these signaling pathways and their regulatory effects on HFSCs have been previously studied, but many mechanisms are still unclear. This article reviews the regulation of hair follicles, HFSCs and related signaling pathways, with the aims of summarizing previous research results, revealing the regulatory mechanisms of HFSC proliferation and differentiation and providing important references and new ideas for treating clinical diseases.
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Affiliation(s)
| | | | - Jia He
- Department of Burn Surgery, The First People’s Hospital of Foshan, Foshan 528000, China
| | - Jingru Wang
- Department of Burn Surgery, The First People’s Hospital of Foshan, Foshan 528000, China
| | - Xiaodong Chen
- Correspondence. Xiaodong Chen, E-mail: ; Ronghua Yang,
| | - Ronghua Yang
- Correspondence. Xiaodong Chen, E-mail: ; Ronghua Yang,
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20
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Xie S, Chen L, Zhang M, Zhang C, Li H. Self-assembled complete hair follicle organoids by coculture of neonatal mouse epidermal cells and dermal cells in Matrigel. ANNALS OF TRANSLATIONAL MEDICINE 2022; 10:767. [PMID: 35965801 PMCID: PMC9372662 DOI: 10.21037/atm-22-3252] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 06/01/2022] [Accepted: 07/08/2022] [Indexed: 02/05/2023]
Abstract
BACKGROUND 3D organoid cultures of hair follicles (HFs) are powerful models that mimic native HF for both in-depth study of HF disease and precision therapy. However, few studies have investigated the complete structure and properties of HF organoids. To investigate and characterize the complete HF organoids self-assembled by coculture of neonatal mouse epidermal cells (MECs) and dermal cells in Matrigel. METHODS Fresh epidermal and dermal cells from newborn mice (n=4) were isolated, and cocultured (1:1 ratio) in Matrigel using DMEM/F12 medium for 1 week. During the culture, an inverted microscope was used to observe the morphology of the 3D constructs. After 1 week, hematoxylin-eosin (HE) and immunofluorescence (IF) staining of HF-related markers (K5, K73, AE13, and K10), HF stem cell markers (K15, CD34, CD49f), skin-derived precursor-related marker (Nestin), and dermal papillae (DP)-specific markers (SOX2 and ALP) was performed in the harvested constructs to identify the HF organoids. RESULTS Epidermal and dermal cells self-assembled into HF organoids comprising an infundibular cyst-like structure, a lower segment-like structure, and a bulb-like structure from tail to root. The HF organoid had multiple, well-defined compartments similar to native anagen HF. Of the three segments, K73 was expressed in the inner root sheath-like layer, AE13 was localized in the hair shaft-like structure, K5, K15, CD34, and CD49f were present in the outer root sheath-like layer, Nestin labeled the connective tissue sheath-like layer, and SOX2 and ALP were expressed in the DP-like structure. Furthermore, K10 and K73 were expressed in the infundibular cyst-like structure. The expression of these molecular proteins was consistent with native anagen HF. CONCLUSIONS The complete HF organoid regenerated in Matrigel has specific compartments and is an excellent model to study HF disease and precision therapy.
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Affiliation(s)
- Sitian Xie
- Department of Plastic Surgery and Burn Center, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China
| | - Liyun Chen
- Department of Plastic Surgery and Burn Center, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China
| | - Mingjun Zhang
- Department of Plastic Surgery and Burn Center, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China
| | - Cuiping Zhang
- Wound Healing and Cell Biology Laboratory, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing, China
| | - Haihong Li
- Department of Plastic Surgery and Burn Center, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China
- Department of Wound Repair, Institute of Wound Repair and Regeneration Medicine, Southern University of Science and Technology Hospital, Southern University of Science and Technology School of Medicine, Shenzhen, China
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21
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Lee J, van der Valk WH, Serdy SA, Deakin C, Kim J, Le AP, Koehler KR. Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells. Nat Protoc 2022; 17:1266-1305. [PMID: 35322210 PMCID: PMC10461778 DOI: 10.1038/s41596-022-00681-y] [Citation(s) in RCA: 28] [Impact Index Per Article: 9.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2021] [Accepted: 01/04/2022] [Indexed: 12/28/2022]
Abstract
Human skin uses millions of hairs and glands distributed across the body surface to function as an external barrier, thermoregulator and stimuli sensor. The large-scale generation of human skin with these appendages would be beneficial, but is challenging. Here, we describe a detailed protocol for generating hair-bearing skin tissue entirely from a homogeneous population of human pluripotent stem cells in a three-dimensional in vitro culture system. Defined culture conditions are used over a 2-week period to induce differentiation of pluripotent stem cells to surface ectoderm and cranial neural crest cells, which give rise to the epidermis and dermis, respectively, in each organoid unit. After 60 d of incubation, the skin organoids produce hair follicles. By day ~130, the skin organoids reach full complexity and contain stratified skin layers, pigmented hair follicles, sebaceous glands, Merkel cells and sensory neurons, recapitulating the cell composition and architecture of fetal skin tissue at week 18 of gestation. Skin organoids can be maintained in culture using this protocol for up to 150 d, enabling the organoids to be used to investigate basic skin biology, model disease and, further, reconstruct or regenerate skin tissue.
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Affiliation(s)
- Jiyoon Lee
- Department of Otolaryngology, Boston Children's Hospital, Boston, MA, USA.
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA.
- Department of Plastic and Oral Surgery, Boston Children's Hospital, Boston, MA, USA.
- Department of Surgery, Harvard Medical School, Boston, MA, USA.
- Department of Otolaryngology-Head and Neck Surgery, Harvard Medical School, Boston, MA, USA.
| | - Wouter H van der Valk
- Department of Otolaryngology, Boston Children's Hospital, Boston, MA, USA
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA
- Department of Otolaryngology-Head and Neck Surgery, Harvard Medical School, Boston, MA, USA
- Department of Otorhinolaryngology and Head & Neck Surgery, Leiden University Medical Center, Leiden, the Netherlands
| | - Sara A Serdy
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA
| | - CiCi Deakin
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA
- Department of Biological Engineering, Wentworth Institute of Technology, Boston, MA, USA
| | - Jin Kim
- Department of Otolaryngology, Boston Children's Hospital, Boston, MA, USA
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA
- Department of Plastic and Oral Surgery, Boston Children's Hospital, Boston, MA, USA
- Department of Otolaryngology-Head and Neck Surgery, Harvard Medical School, Boston, MA, USA
| | - Anh Phuong Le
- Department of Otolaryngology, Boston Children's Hospital, Boston, MA, USA
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA
- Department of Plastic and Oral Surgery, Boston Children's Hospital, Boston, MA, USA
- Department of Otolaryngology-Head and Neck Surgery, Harvard Medical School, Boston, MA, USA
| | - Karl R Koehler
- Department of Otolaryngology, Boston Children's Hospital, Boston, MA, USA.
- F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA, USA.
- Department of Plastic and Oral Surgery, Boston Children's Hospital, Boston, MA, USA.
- Department of Otolaryngology-Head and Neck Surgery, Harvard Medical School, Boston, MA, USA.
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22
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Direct Reprograming of Mouse Fibroblasts into Dermal Papilla Cells via Small Molecules. Int J Mol Sci 2022; 23:ijms23084213. [PMID: 35457029 PMCID: PMC9030401 DOI: 10.3390/ijms23084213] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2022] [Revised: 03/29/2022] [Accepted: 04/07/2022] [Indexed: 01/27/2023] Open
Abstract
The reprogramming of somatic fibroblasts into alternative cell linages could provide a promising source of cells for regenerative medicine and cell therapy. However, the direct conversion of fibroblasts into other functional cell types is still challenging. In this study, we show that dermal-papilla-cell-like cells (DPC-LCs) can be generated by treating fibroblasts, including L929 mouse fibroblast cell lines and somatic mouse fibroblasts, with small molecules. Based on alkaline phosphatase activity and other molecular markers, different compounds or their combinations are needed for converting the two different fibroblasts into DPC-LCs. Notably, we found that TTNPB alone can efficiently convert primary adult mouse fibroblasts into DPC-LCs. DPC-LCs generated from mouse fibroblasts showed a stronger hair-inducing capacity. Transcriptome analysis reveals that expression of genes associated with a hair-inducing capacity are increased in DPC-LCs. This pharmacological approach to generating functional dermal papilla cells may have many important implications for hair follicle regeneration and hair loss therapy.
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23
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Inhibition of class I HDACs preserves hair follicle inductivity in postnatal dermal cells. Sci Rep 2021; 11:24056. [PMID: 34911993 PMCID: PMC8674223 DOI: 10.1038/s41598-021-03508-0] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/24/2021] [Accepted: 12/03/2021] [Indexed: 11/09/2022] Open
Abstract
Induction of new hair follicles (HFs) may be an ultimate treatment goal for alopecia; however, functional cells with HF inductivity must be expanded in bulk for clinical use. In vitro culture conditions are completely different from the in vivo microenvironment. Although fetal and postnatal dermal cells (DCs) have the potential to induce HFs, they rapidly lose this HF inductivity during culture, accompanied by a drastic change in gene expression. This suggests that epigenetic regulation may be involved. Of the various histone deacetylases (HDACs), Class I HDACs are noteworthy because they are ubiquitously expressed and have the strongest deacetylase activity. This study revealed that DCs from postnatal mice rapidly lose HF inductivity and that this reduction is accompanied by a significant decrease in histone H3 acetylation. However, MS-275, an inhibitor of class I HDACs, preserves HF inductivity in DCs during culture, increasing alkaline phosphatase activity and upregulating HF inductive genes such as BMP4, HEY1, and WIF1. In addition, the inhibition of class I HDACs activates the Wnt signaling pathway, the most well-described molecular pathway in HF development, via increased histone H3 acetylation within the promoter region of the Wnt transcription factor LEF1. Our results suggest that class I HDACs could be a potential target for the neogenesis of HFs.
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24
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Sun C, Hu SH, Dong BQ, Jiang S, Miao F, Lei TC. Metformin Promotes the Hair-Inductive Activity of Three-Dimensional Aggregates of Epidermal and Dermal Cells Self-Assembled In Vitro. Skin Pharmacol Physiol 2021; 35:137-147. [PMID: 34883492 DOI: 10.1159/000521400] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/22/2021] [Accepted: 12/07/2021] [Indexed: 11/19/2022]
Abstract
INTRODUCTION Although it has been reported that the anti-diabetic drug metformin has multiple extra-hypoglycemic activities, such as anti-oxidation, anti-aging and even anti-tumor, topical metformin also can induce hair regeneration, but the precise mechanism involved in that process is still unclear. OBJECTIVES To assess the effect of metformin on hair growth in a mouse hair follicle reconstitution model generated by in vitro self-assembled three-dimensional aggregates of epidermal and dermal cells (3D aggregates). METHODS Epidermal cells and dermal cells were isolated and cultured from the mouse skin of fifty C57BL/6 mouse pups (1-day-old). For tracing the distribution of dermal cells during the self-assembly process of 3D aggregates, the dermal cells were labeled with Vybrant Dil cell-labelling solution and mixed with epidermal cells at 1:1 ratio. Formed 3D aggregates were treated with 10 mM metformin and then were grafted into recipient BALB/c nude mice. The biomarkers (HGF, CD133, ALP, β-catenin and SOX2) associated with the hair-inductive activity of dermal cells were detected in the grafted skin tissues and in cultured 3D aggregates treated with metformin using immunofluorescent staining, quantitative real-time RT-PCR (qRT-PCR), and western blotting. Furthermore, the expression levels of CD133 were also examined in dermal cells with different passage numbers using qRT-PCR and western blotting. RESULTS Metformin directly stimulates the activity of alkaline phosphatase (ALP) of cultured 3D aggregates, upregulates both the protein and mRNA expression levels of molecular markers (HGF, CD133, ALP, β-catenin and SOX2) and improves the survival rate of reconstituted hair follicles. Moreover, we also found that metformin increases the expression of CD133 in dermal cells thus maintaining their trichogenic capacity that would normally be lost by serial subculture. CONCLUSIONS These results suggest that metformin can promote hair follicle regeneration in vitro through up-regulation of the hair inductive capability of dermal cells, warranting further evaluation in the clinical treatment of male or female pattern hair loss.
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Affiliation(s)
- Chao Sun
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China,
| | - Shuang-Hai Hu
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China
| | - Bing-Qi Dong
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China
| | - Shan Jiang
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China
| | - Fang Miao
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China
| | - Tie-Chi Lei
- Department of Dermatology, Renmin Hospital of Wuhan University, Wuhan, China
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25
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Abstract
SUMMARY The advent of pluripotent stem cells following the discovery of Shinya Yamanaka (2012 Nobel prize in Medicine) brought about a regenerative medicine approach to virtually every human condition including hair loss. It is now possible to reprogram somatic cells (eg, blood or skin cells) from a person experiencing hair loss to generate autologous induced pluripotent stem cells (iPSCs), which could be amplified and cryopreserved. Subsequently, these iPSCs could be differentiated into various cell types such as dermal papilla cells, epithelial cells, melanocytes, and other cell types constituting functional hair follicle. Transplantation of human iPSC-derived folliculogenic cells into the nude mice has successfully generated xenografts with hair outgrowth. Because iPSCs provide a virtually unlimited source of folliculogenic cells for de novo formation of hair follicles, this approach has major advantages over current surgical hair restoration procedures, which merely redistribute existing hair follicles from one part of the sculp to another. Combined with robotics and automation of the transplantation process, this novel regenerative medicine approach is well poised to make hair restoration a routine procedure affordable for everybody who can benefit from it.
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26
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Kang D, Liu Z, Qian C, Huang J, Zhou Y, Mao X, Qu Q, Liu B, Wang J, Wang Y, Hu Z, Huang W, Miao Y. A three-dimensional bioprinting technique, based on a gelatin/alginate hydrogel, for the tissue engineering of hair follicle reconstruction. Int J Biol Macromol 2021:S0141-8130(21)01927-9. [PMID: 34509522 DOI: 10.1016/j.ijbiomac.2021.09.014] [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: 06/03/2021] [Revised: 08/31/2021] [Accepted: 09/02/2021] [Indexed: 12/17/2022]
Abstract
Hair loss remains a challenging clinical problem that influences the quality of life. Three-dimensional (3D) bioprinting has become a valuable tool for fabricating tissue constructs for transplantation and other biomedical applications. Although some simple organs, such as skin and cartilage, have been successfully simulated, it remains challenging to make hair follicles (HFs), which are highly complex organs. The tissue engineering of human HFs has been a long-standing challenge, and progress with this has lagged behind that with other lab-grown tissues. This is principally due to a lack of availability of a platform that can successfully recapitulate the microenvironmental cues required to maintain the requisite cellular interactions for hair neogenesis. In this study, we used a 3D bioprinting technique based on a gelatin/alginate hydrogel to construct a multilayer composite scaffold with cuticular and corium layers to simulate the microenvironment of dermal papilla cells (DPCs) in the human body. This new approach permits the controllable formation of self-aggregating spheroids of DPCs in a physiologically relevant extracellular matrix and the initiation of epidermal-mesenchymal interactions, which results in HF formation in vivo. In conclusion, our 3D-bioprinted multilayer composite scaffold prepared using a gelatin/alginate hydrogel provides a suitable 3D microenvironment for DPCs to induce HF formation. The ability to regenerate entire HFs should have a significant impact on the medical management of hair loss. This method may also have critical applications for skin tissue engineering, with its appendages, for other purposes.
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Affiliation(s)
- Deni Kang
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Zhen Liu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Chuanmu Qian
- Department of Anesthesiology, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong 510317, China
| | - Junfei Huang
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Yi Zhou
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Xiaoyan Mao
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Qian Qu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Bingcheng Liu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Jin Wang
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Yilin Wang
- Department of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China
| | - Zhiqi Hu
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China.
| | - Wenhua Huang
- Department of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China.
| | - Yong Miao
- Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China.
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27
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He J, Zhang X, Xia X, Han M, Li F, Li C, Li Y, Gao D. Organoid technology for tissue engineering. J Mol Cell Biol 2021; 12:569-579. [PMID: 32249317 PMCID: PMC7683016 DOI: 10.1093/jmcb/mjaa012] [Citation(s) in RCA: 35] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2019] [Revised: 01/11/2020] [Accepted: 02/04/2020] [Indexed: 12/18/2022] Open
Abstract
For centuries, attempts have been continuously made to artificially reconstitute counterparts of in vivo organs from their tissues or cells. Only in the recent decade has organoid technology as a whole technological field systematically emerged and been shown to play important roles in tissue engineering. Based on their self-organizing capacities, stem cells of versatile organs, both harvested and induced, can form 3D structures that are structurally and functionally similar to their in vivo counterparts. These organoid models provide a powerful platform for elucidating the development mechanisms, modeling diseases, and screening drug candidates. In this review, we will summarize the advances of this technology for generating various organoids of tissues from the three germ layers and discuss their drawbacks and prospects for tissue engineering.
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Affiliation(s)
- Juan He
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Xiaoyu Zhang
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Xinyi Xia
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Ming Han
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Fei Li
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Chunfeng Li
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Yunguang Li
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
| | - Dong Gao
- State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.,Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China
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28
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Fukuyama M, Tsukashima A, Kimishima M, Yamazaki Y, Okano H, Ohyama M. Human iPS Cell-Derived Cell Aggregates Exhibited Dermal Papilla Cell Properties in in vitro Three-Dimensional Assemblage Mimicking Hair Follicle Structures. Front Cell Dev Biol 2021; 9:590333. [PMID: 34409023 PMCID: PMC8365839 DOI: 10.3389/fcell.2021.590333] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/01/2020] [Accepted: 07/07/2021] [Indexed: 12/11/2022] Open
Abstract
Current approaches for human hair follicle (HF) regeneration mostly adopt cell-autonomous tissue reassembly in a permissive murine intracorporeal environment. This, together with the limitation in human-derived trichogenic starting materials, potentially hinders the bioengineering of human HF structures, especially for the drug discovery and treatment of hair loss disorders. In this study, we attempted to reproduce the anatomical relationship between an epithelial main body and the dermal papilla (DP) within HF in vitro by three-dimensionally assembling columnarly molded human keratinocytes (KCs) and the aggregates of DP cells and evaluated how HF characteristics were reproduced in the constructs. The replaceability of human-induced pluripotent stem cell (hiPSC)-derived DP substitutes was assessed using the aforementioned reconstruction assay. Human DP cell aggregates were embedded into Matrigel as a cluster. Subsequently, highly condensed human KCs were cylindrically injected onto DP spheroids. After 2-week culture, the structures visually mimicking HFs were obtained. KC-DP constructs partially reproduced HF microanatomy and demonstrated differential keratin (KRT) expression pattern in HFs: KRT14 in the outermost part and KRT13, KRT17, and KRT40, respectively, in the inner portion of the main body. KC-DP constructs tended to upregulate HF-related genes, KRT25, KRT33A, KRT82, WNT5A, and LEF1. Next, DP substitutes were prepared by exposing hiPSC-derived mesenchymal cells to retinoic acid and subsequently to WNT, BMP, and FGF signal activators, followed by cell aggregation. The resultant hiPSC-derived DP substitutes (iDPs) were combined with KCs in the invented assay. KC-iDP constructs morphologically resemble KC-DP constructs and analogously mimicked KRT expression pattern in HF. iDP in the constructs expressed DP-related markers, such as vimentin and versican. Intriguingly, KC-iDP constructs more intensely expressed KRT33A, KRT82, and LEF1, which were stepwisely upregulated by the addition of WNT ligand and the mixture of WNT, SHH, and EDA signaling activators, supporting the idea that iDP exhibited biological properties analogous to DP cell aggregates in the constructs in vitro. These preliminary findings suggested the possibility of regenerating DP equivalents with in vitro hair-inductive capacity using hiPSC-derived cell composites, which potentially reduce the necessity of human tissue-derived trichogenic cell subset and eventually allow xeno-free bioengineering of human HFs.
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Affiliation(s)
- Masahiro Fukuyama
- Department of Dermatology, Kyorin University Faculty of Medicine, Tokyo, Japan
| | - Aki Tsukashima
- Department of Dermatology, Kyorin University Faculty of Medicine, Tokyo, Japan
| | - Momoko Kimishima
- Department of Dermatology, Kyorin University Faculty of Medicine, Tokyo, Japan
| | - Yoshimi Yamazaki
- Department of Dermatology, Kyorin University Faculty of Medicine, Tokyo, Japan
| | - Hideyuki Okano
- Department of Physiology, Keio University School of Medicine, Tokyo, Japan
| | - Manabu Ohyama
- Department of Dermatology, Kyorin University Faculty of Medicine, Tokyo, Japan
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29
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A carbazole compound, 9-ethyl-9H-carbazole-3-carbaldehyde, plays an antitumor function through reactivation of the p53 pathway in human melanoma cells. Cell Death Dis 2021; 12:591. [PMID: 34103468 PMCID: PMC8187445 DOI: 10.1038/s41419-021-03867-6] [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: 02/04/2021] [Revised: 05/24/2021] [Accepted: 05/24/2021] [Indexed: 11/30/2022]
Abstract
p53, the major tumor suppressor, is frequently mutated in many cancers, and up to 84% of human melanomas harbor wild-type p53, which is considered to be an ideal target for melanoma therapy. Here, we evaluated the antitumor activity of a carbazole derivative, 9-ethyl-9H-carbazole-3-carbaldehyde (ECCA), on melanoma cells. ECCA had a selectively strong inhibitory activity against the growth of BRAF-mutated and BRAF-wild-type melanoma cells but had little effect on normal human primary melanocytes. ECCA inhibited melanoma cell growth by increasing cell apoptosis, which was associated with the upregulation of caspase activities and was significantly abrogated by the addition of a caspase inhibitor. In vivo assays confirmed that ECCA suppressed melanoma growth by enhancing cell apoptosis and reducing cell proliferation, and importantly ECCA did not have any evident toxic effects on normal tissues. RNA-Seq analysis identified several pathways related to cell apoptosis that were affected by ECCA, notably, activation of the p53 signaling pathway. Biochemical assays demonstrated that ECCA enhanced the phosphorylation of p53 at Ser15 in melanoma cells harboring wild-type p53, and importantly, the knockdown or deletion of p53 in those cells counteracted the ECCA-induced apoptosis, as well as senescence. Further investigations revealed that ECCA enhanced the phosphorylation of p38-MAPK and c-Jun N-terminal kinase (JNK), and treatment with either a p38-MAPK or a JNK inhibitor rescued the cell growth inhibition elicited by ECCA, which depended on the expression of the p53 gene. Finally, the combination of ECCA with a BRAF inhibitor significantly enhanced the growth inhibition of melanoma cells. In summary, our study demonstrates that the carbazole derivative, ECCA, induces melanoma cell apoptosis and senescence through the activation of p53 to significantly and selectively suppress the growth of melanoma cells without affecting normal human melanocytes, suggesting its potential to develop a new drug for melanoma therapy.
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30
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Ibrahim MR, Medhat W, El-Fakahany H, Abdel-Raouf H, Snyder EY. The Developmental & Molecular Requirements for Ensuring that Human Pluripotent Stem Cell-Derived Hair Follicle Bulge Stem Cells Have Acquired Competence for Hair Follicle Generation Following Transplantation. Cell Transplant 2021; 30:9636897211014820. [PMID: 34053245 PMCID: PMC8182633 DOI: 10.1177/09636897211014820] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023] Open
Abstract
When using human induced pluripotent stem cells (hiPSCs) to achieve hair follicle (HF) replacement, we found it best to emulate the earliest fundamental developmental processes of gastrulation, ectodermal lineage commitment, and dermogenesis. Viewing hiPSCs as a model of the epiblast, we exploited insights from mapping the dynamic up- and down-regulation of the developmental molecules that determine HF lineage in order to ascertain the precise differentiation stage and molecular requirements for grafting HF-generating progenitors. To yield an integrin-dependent lineage like the HF in vivo, we show that hiPSC derivatives should co-express, just prior to transplantation, the following combination of markers: integrins α6 and β1 and the glycoprotein CD200 on their surface; and, intracellularly, the epithelial marker keratin 18 and the hair follicle bulge stem cell (HFBSC)-defining molecules transcription factor P63 and the keratins 15 and 19. If the degree of trichogenic responsiveness indicated by the presence of these molecules is not achieved (they peak on Days 11-18 of the protocol), HF generation is not possible. Conversely, if differentiation of the cells is allowed to proceed beyond the transient intermediate progenitor state represented by the HFBSC, and instead cascades to their becoming keratin 14+ keratin 5+ CD200– keratinocytes (Day 25), HF generation is equally impossible. We make the developmental case for transplanting at Day 16-18 of differentiation—the point at which the hiPSCs have lost pluripotency, have attained optimal expression of HFBSC markers, have not yet experienced downregulation of key integrins and surface glycoproteins, have not yet started expressing keratinocyte-associated molecules, and have sufficient proliferative capacity to allow a well-populated graft. This panel of markers may be used for isolating (by cytometry) HF-generating derivatives away from cell types unsuited for this therapy as well as for identifying trichogenic drugs.
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Affiliation(s)
- Michel R Ibrahim
- Department of Dermatology, STD's and Andrology, Faculty of Medicine, Minia University, Al-Minya, Egypt.,Center for Stem Cells & Regenerative Medicine, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA.,Sanford Consortium for Regenerative Medicine, La Jolla, CA, USA
| | - Walid Medhat
- Department of Dermatology, STD's and Andrology, Faculty of Medicine, Minia University, Al-Minya, Egypt
| | - Hasan El-Fakahany
- Department of Dermatology, STD's and Andrology, Faculty of Medicine, Minia University, Al-Minya, Egypt
| | - Hamza Abdel-Raouf
- Department of Dermatology, STD's and Andrology, Faculty of Medicine, Minia University, Al-Minya, Egypt
| | - Evan Y Snyder
- Center for Stem Cells & Regenerative Medicine, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA.,Sanford Consortium for Regenerative Medicine, La Jolla, CA, USA.,Department of Pediatrics, University of California-San Diego, La Jolla, CA, USA
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31
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Žnidarič M, Žurga ŽM, Maver U. Design of In Vitro Hair Follicles for Different Applications in the Treatment of Alopecia-A Review. Biomedicines 2021; 9:biomedicines9040435. [PMID: 33923738 PMCID: PMC8072628 DOI: 10.3390/biomedicines9040435] [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: 03/22/2021] [Revised: 04/08/2021] [Accepted: 04/13/2021] [Indexed: 12/19/2022] Open
Abstract
The hair research field has seen great improvement in recent decades, with in vitro hair follicle (HF) models being extensively developed. However, due to the cellular complexity and number of various molecular interactions that must be coordinated, a fully functional in vitro model of HFs remains elusive. The most common bioengineering approach to grow HFs in vitro is to manipulate their features on cellular and molecular levels, with dermal papilla cells being the main focus. In this study, we focus on providing a better understanding of HFs in general and how they behave in vitro. The first part of the review presents skin morphology with an emphasis on HFs and hair loss. The remainder of the paper evaluates cells, materials, and methods of in vitro growth of HFs. Lastly, in vitro models and assays for evaluating the effects of active compounds on alopecia and hair growth are presented, with the final emphasis on applications of in vitro HFs in hair transplantation. Since the growth of in vitro HFs is a complicated procedure, there is still a great number of unanswered questions aimed at understanding the long-term cycling of HFs without losing inductivity. Incorporating other regions of HFs that lead to the successful formation of different hair classes remains a difficult challenge.
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32
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Nakajima R, Tate Y, Yan L, Kageyama T, Fukuda J. Impact of adipose-derived stem cells on engineering hair follicle germ-like tissue grafts for hair regenerative medicine. J Biosci Bioeng 2021; 131:679-685. [PMID: 33678531 DOI: 10.1016/j.jbiosc.2021.02.001] [Citation(s) in RCA: 13] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2020] [Revised: 02/01/2021] [Accepted: 02/04/2021] [Indexed: 12/11/2022]
Abstract
Hair regenerative medicine has emerged as a promising treatment strategy for severe hair loss, such as end-stage androgenetic alopecia. Various approaches to engineering three-dimensional tissue grafts have been explored since they drive the ability to regenerate hair follicles when transplanted. In the present study, we demonstrated the assembly of human adipose-derived stem cells (hASCs) into hair follicle germ (HFG)-like aggregates for de novo hair regeneration. We mixed human dermal papilla cells (hDPCs), murine embryonic epithelial cells, and hASCs in suspension, and allowed them to form aggregates. During three days of culture, cells initially formed a single aggregate with a random distribution of the three cell types, but the epithelial and dermal papilla cells subsequently separated from each other and formed a dumbbell-shaped HFG, with hASCs localized on the hDPC aggregate side. The involvement of hASCs significantly increased gene expression associated with hair morphogenesis compared to HFGs without hASCs. The self-organization of the three cell types was observed in our scalable lab-made chip device. HFGs containing hASCs efficiently generated hair shafts upon transplantation to nude mice, while only a few shafts were generated with HFGs without hASCs. This approach may be a promising strategy for fabricating tissue grafts for hair regenerative medicine.
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Affiliation(s)
- Rikuma Nakajima
- Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Yoshiki Tate
- Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Lei Yan
- Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan
| | - Tatsuto Kageyama
- Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan; Kanagawa Institute of Industrial Science and Technology, 3-2-1 Sakado Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan; Japan Science and Technology Agency (JST)-PRESTO, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
| | - Junji Fukuda
- Faculty of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama, Kanagawa 240-8501, Japan; Kanagawa Institute of Industrial Science and Technology, 3-2-1 Sakado Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
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Tan JJ, Nguyen DV, Common JE, Wu C, Ho PC, Kang L. Investigating PEGDA and GelMA Microgel Models for Sustained 3D Heterotypic Dermal Papilla and Keratinocyte Co-Cultures. Int J Mol Sci 2021; 22:2143. [PMID: 33670029 PMCID: PMC7926670 DOI: 10.3390/ijms22042143] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/20/2021] [Revised: 02/13/2021] [Accepted: 02/17/2021] [Indexed: 02/05/2023] Open
Abstract
Hair follicle morphogenesis is heavily dependent on reciprocal, sequential, and epithelial-mesenchymal interaction (EMI) between epidermal stem cells and the specialized cells of the underlying mesenchyme, which aggregate to form the dermal condensate (DC) and will later become the dermal papilla (DP). Similar models were developed with a co-culture of keratinocytes and DP cells. Previous studies have demonstrated that co-culture with keratinocytes maintains the in vivo characteristics of the DP. However, it is often challenging to develop three-dimensional (3D) DP and keratinocyte co-culture models for long term in vitro studies, due to the poor intercellular adherence between keratinocytes. Keratinocytes exhibit exfoliative behavior, and the integrity of the DP and keratinocyte co-cultured spheroids cannot be maintained over prolonged culture. Short durations of culture are unable to sufficiently allow the differentiation and re-programming of the keratinocytes into hair follicular fate by the DP. In this study, we explored a microgel array approach fabricated with two different hydrogel systems. Using poly (ethylene glycol) diacrylate (PEGDA) and gelatin methacrylate (GelMA), we compare their effects on maintaining the integrity of the cultures and their expression of important genes responsible for hair follicle morphogenesis, namely Wnt10A, Wnt10B, and Shh, over prolonged duration. We discovered that low attachment surfaces such as PEGDA result in the exfoliation of keratinocytes and were not suitable for long-term culture. GelMA, on the hand, was able to sustain the integrity of co-cultures and showed higher expression of the morphogens overtime.
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Affiliation(s)
- Justin J.Y. Tan
- Department of Pharmacy, National University of Singapore, Lower Kent Ridge Road, 18 Science Drive 4, Singapore 117543, Singapore; (J.J.Y.T.); (P.C.L.H.)
| | - Duc-Viet Nguyen
- Nusmetics Pte. Ltd., i4 Building, 3 Research Link, Singapore 117602, Singapore;
| | - John E. Common
- Skin Research Institute of Singapore, Immunos, 8A Biomedical Grove, Singapore 138648, Singapore;
| | - Chunyong Wu
- Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China;
| | - Paul C.L. Ho
- Department of Pharmacy, National University of Singapore, Lower Kent Ridge Road, 18 Science Drive 4, Singapore 117543, Singapore; (J.J.Y.T.); (P.C.L.H.)
| | - Lifeng Kang
- School of Pharmacy, University of Sydney, Pharmacy and Bank Building A15, Sydney, NSW 2006, Australia
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Atwood SX, Plikus MV. Fostering a healthy culture: Biological relevance of in vitro and ex vivo skin models. Exp Dermatol 2021; 30:298-303. [PMID: 33565670 DOI: 10.1111/exd.14296] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Affiliation(s)
- Scott X Atwood
- Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA.,Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA.,NSF-Simons Center for Multiscale Cell Fate Research, University of California, Irvine, Irvine, CA, USA
| | - Maksim V Plikus
- Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA.,Center for Complex Biological Systems, University of California, Irvine, Irvine, CA, USA.,NSF-Simons Center for Multiscale Cell Fate Research, University of California, Irvine, Irvine, CA, USA.,Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, USA
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Qiu W, Gu PR, Chuong CM, Lei M. Skin Cyst: A Pathological Dead-End With a New Twist of Morphogenetic Potentials in Organoid Cultures. Front Cell Dev Biol 2021; 8:628114. [PMID: 33511139 PMCID: PMC7835531 DOI: 10.3389/fcell.2020.628114] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2020] [Accepted: 12/17/2020] [Indexed: 01/07/2023] Open
Abstract
A cyst is a closed sac-like structure in which cyst walls wrap certain contents typically including air, fluid, lipid, mucous, or keratin. Cyst cells can retain multipotency to regenerate complex tissue architectures, or to differentiate. Cysts can form in and outside the skin due to genetic problems, errors in embryonic development, cellular defects, chronic inflammation, infections, blockages of ducts, parasites, and injuries. Multiple types of skin cysts have been identified with different cellular origins, with a common structure including the outside cyst wall engulfs differentiated suprabasal layers and keratins. The skin cyst is usually used as a sign in pathological diagnosis. Large or surfaced skin cysts affect patients' appearance and may cause the dysfunction or accompanying diseases of adjacent tissues. Skin cysts form as a result of the degradation of skin epithelium and appendages, retaining certain characteristics of multipotency. Surprisingly, recent organoid cultures show the formation of cyst configuration as a transient state toward more morphogenetic possibility. These results suggest, if we can learn more about the molecular circuits controlling upstream and downstream cellular events in cyst formation, we may be able to engineer stem cell cultures toward the phenotypes we wish to achieve. For pathological conditions in patients, we speculate it may also be possible to guide the cyst to differentiate or de-differentiate to generate structures more akin to normal architecture and compatible with skin homeostasis.
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Affiliation(s)
- Weiming Qiu
- Department of Dermatology, General Hospital of Central Theater Command of Chinese People’s Liberation Army, Wuhan, China
| | - Pei-Rong Gu
- Integrative Stem Cell Center, China Medical University Hospital, China Medical University, Taichung, Taiwan
| | - Cheng-Ming Chuong
- Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, United States
| | - Mingxing Lei
- Integrative Stem Cell Center, China Medical University Hospital, China Medical University, Taichung, Taiwan
- “111” Project Laboratory of Biomechanics and Tissue Repair, Key Laboratory of Biorheological Science and Technology of the Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China
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36
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Bak SS, Park JM, Oh JW, Kim JC, Kim MK, Sung YK. Knockdown of FOXA2 Impairs Hair-Inductive Activity of Cultured Human Follicular Keratinocytes. Front Cell Dev Biol 2020; 8:575382. [PMID: 33117803 PMCID: PMC7578224 DOI: 10.3389/fcell.2020.575382] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/23/2020] [Accepted: 09/17/2020] [Indexed: 01/12/2023] Open
Abstract
Reciprocal interactions between hair-inductive dermal cells and epidermal cells are essential for de novo genesis of hair follicles. Recent studies have shown that outer root sheath (ORS) follicular keratinocytes can be expanded in vitro, but the cultured cells often lose receptivity to hair-inducing dermal signals. In this study, we first investigated whether the hair-inductive activity (trichogenicity) of cultured human ORS follicular keratinocytes was correlated with the cultivation period. ORS follicular keratinocytes from the scalp were cultured for 3, 4, 5, or 6 weeks and were then implanted into nude mice along with freshly isolated neonatal mouse dermal cells. We observed that the trichogenicity of the implanted ORS cells was inversely correlated with their cultivation period. These initial findings prompted us to investigate the differentially expressed genes between the short-term (20 days) and long-term (42 days) cultured ORS cells, trichogenic and non-trichogenic, respectively, by microarray analysis. We found that forkhead box protein A2 (FOXA2) was the most up-regulated transcription factor in the trichogenic ORS cells. Thus, we investigated whether the trichogenicity of the cells was affected by FOXA2 expression. We found a significant decrease in the number of induced hair follicles when the ORS cells were transfected with a FOXA2 small interfering RNA versus control small interfering RNA. Taken together, our data strongly suggest that FOXA2 significantly influences the trichogenicity of human ORS cells.
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Affiliation(s)
- Soon-Sun Bak
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu, South Korea
| | - Jung Min Park
- Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, South Korea.,Clinical Omics Institute, Kyungpook National University, Daegu, South Korea
| | - Ji Won Oh
- Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, South Korea.,Clinical Omics Institute, Kyungpook National University, Daegu, South Korea.,Hair Transplantation Center, Kyungpook National University Hospital, Daegu, South Korea
| | - Jung Chul Kim
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu, South Korea.,Hair Transplantation Center, Kyungpook National University Hospital, Daegu, South Korea
| | - Moon Kyu Kim
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu, South Korea.,Hair Transplantation Center, Kyungpook National University Hospital, Daegu, South Korea
| | - Young Kwan Sung
- Department of Immunology, School of Medicine, Kyungpook National University, Daegu, South Korea
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37
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Abbasi S, Sinha S, Labit E, Rosin NL, Yoon G, Rahmani W, Jaffer A, Sharma N, Hagner A, Shah P, Arora R, Yoon J, Islam A, Uchida A, Chang CK, Stratton JA, Scott RW, Rossi FMV, Underhill TM, Biernaskie J. Distinct Regulatory Programs Control the Latent Regenerative Potential of Dermal Fibroblasts during Wound Healing. Cell Stem Cell 2020; 27:396-412.e6. [PMID: 32755548 DOI: 10.1016/j.stem.2020.07.008] [Citation(s) in RCA: 124] [Impact Index Per Article: 24.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2020] [Revised: 05/25/2020] [Accepted: 07/09/2020] [Indexed: 01/12/2023]
Abstract
Dermal fibroblasts exhibit considerable heterogeneity during homeostasis and in response to injury. Defining lineage origins of reparative fibroblasts and regulatory programs that drive fibrosis or, conversely, promote regeneration will be essential for improving healing outcomes. Using complementary fate-mapping approaches, we show that hair follicle mesenchymal progenitors make limited contributions to wound repair. In contrast, extrafollicular progenitors marked by the quiescence-associated factor Hic1 generated the bulk of reparative fibroblasts and exhibited functional divergence, mediating regeneration in the center of the wound neodermis and scar formation in the periphery. Single-cell RNA-seq revealed unique transcriptional, regulatory, and epithelial-mesenchymal crosstalk signatures that enabled mesenchymal competence for regeneration. Integration with scATAC-seq highlighted changes in chromatin accessibility within regeneration-associated loci. Finally, pharmacological modulation of RUNX1 and retinoic acid signaling or genetic deletion of Hic1 within wound-activated fibroblasts was sufficient to modulate healing outcomes, suggesting that reparative fibroblasts have latent but modifiable regenerative capacity.
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Affiliation(s)
- Sepideh Abbasi
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Sarthak Sinha
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Elodie Labit
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Nicole L Rosin
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Grace Yoon
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Waleed Rahmani
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Arzina Jaffer
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Nilesh Sharma
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Andrew Hagner
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Prajay Shah
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Rohit Arora
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Jessica Yoon
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Anowara Islam
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Aya Uchida
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - Chih Kai Chang
- Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada
| | - Jo Anne Stratton
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
| | - R Wilder Scott
- Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada
| | - Fabio M V Rossi
- Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada
| | - T Michael Underhill
- Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada
| | - Jeff Biernaskie
- Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada; Alberta Children's Hospital Research Institute, University of Calgary, Calgary, AB T2N 4N1, Canada; Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada.
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38
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Li X, Ye Y, Liu X, Bai L, Zhao P, Bai W, Zhang M. Low-frequency electromagnetic fields promote hair follicles regeneration by injection a mixture of epidermal stem cells and dermal papilla cells. Electromagn Biol Med 2020; 39:251-256. [PMID: 32727226 DOI: 10.1080/15368378.2020.1793165] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Abstract
The bioeffects of low-frequency electromagnetic fields (EMF) on a bio-engineered hair follicle generation had not been fully elucidated. This present study was designed to evaluat the therapeutically effective of low frequency EMF on hair follicles regeneration. In this experiment, epidermal stem cells (ESCs) and dermal papilla (DP) cells were isolated and culture-expanded. Then the mixture containing of ESCs and DP cells was implanted into the epidermal layer or corium layer of nude mice. Those mice were divided at random into the control group and EMF group, 7 days or 14 days later, the skin specimens were harvested to assess for hair regeneration or a bio-engineered skin formation using H&E staining. After injection of the mixture into the epidermal layer of nude mice for 14 days, H&E staining showed that the new hair formed the correct structure comprising hair matrix, hair shaft, and inner root sheath, outer root sheath, and DP. Comparing to the control, the hair follicles erupted at a higher density in the EMF group. When the mixture was implanted into the corium layer for 7 days, comparing with the characteristics of new hair follicles in the control group, H&E staining also showed the mixture induced to form 4 ~ 6 epidermal layers with a higher density of hair follicle like-structures in the bioengineered epithelial layers after EMF exposure. Our results suggested that the injection of a mixture of ESCs and DP cells in combination with EMF exposure facilitated the induction of hair follicle regeneration and a bioengineered skin formation with hair follicle-like structures.
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Affiliation(s)
- Xinping Li
- Department of Physical Medicine and Rehabilitation, Guangdong Geriatric Institute, Guangdong Academy of Medical Sciences & Guangdong Provincial People's Hospital , Guangzhou, China
| | - Yan Ye
- Department of Physical Medicine and Rehabilitation, The Second People' Hospital of Foshan , Foshan, China
| | - Xiaohan Liu
- Department of Physical Medicine and Rehabilitation, The Fifth Affiliated Hospital of Sun Yat-sen University , Zhuhai, China
| | - Liming Bai
- Department of Physical Medicine and Rehabilitation, Guangdong Geriatric Institute, Guangdong Academy of Medical Sciences & Guangdong Provincial People's Hospital , Guangzhou, China
| | - Pin Zhao
- Huayin Laboratory, Southern Medical University , Guangzhou, China
| | - Wenfang Bai
- Department of Physical Medicine and Rehabilitation, Guangdong Geriatric Institute, Guangdong Academy of Medical Sciences & Guangdong Provincial People's Hospital , Guangzhou, China
| | - Mingsheng Zhang
- Department of Physical Medicine and Rehabilitation, Guangdong Geriatric Institute, Guangdong Academy of Medical Sciences & Guangdong Provincial People's Hospital , Guangzhou, China
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Kanayama K, Takada H, Saito N, Kato H, Kinoshita K, Shirado T, Mashiko T, Asahi R, Mori M, Tashiro K, Sunaga A, Kurisaki A, Yoshizato K, Yoshimura K. Hair Regeneration Potential of Human Dermal Sheath Cells Cultured Under Physiological Oxygen. Tissue Eng Part A 2020; 26:1147-1157. [PMID: 32408803 DOI: 10.1089/ten.tea.2019.0329] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022] Open
Abstract
We investigated the effect of oxygen tension on the proliferation and hair-inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were separately obtained from human hair follicles and each cultured under atmospheric/hyperoxic (20% O2), physiological/normoxic (6% O2), or hypoxic (1% O2) conditions. Proliferation of DPCs and DSCs was highest under normoxia. Compared with hyperoxia, hypoxia inhibited proliferation of DPCs, but enhanced that of DSCs. In DPCs, hypoxia downregulated the expression of hair-inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN. In DSCs, both normoxia and hypoxia upregulated SOX2 expression, whereas hypoxia downregulated BMP4 expression. Microarray analysis revealed that normoxia increased the expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, compared with hyperoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In this experiment, normoxia resulted in the most efficient induction of DPC hair follicles, whereas hypoxia caused the most efficient induction and maturation of DSC hair follicles. These results suggest that application of physiological/hypoxic oxygen tension to cultured human DSCs enhances proliferation and maintenance of hair inductivity for skin engineering and clinical applications. Impact statement Dermal sheath cells (DSCs) and dermal papilla cells (DPCs) are useful cell sources for cell-based regenerative therapy. This is the first report to describe that low-oxygen conditions are better for DSCs. Normoxic and hypoxic culture of DSCs is beneficial for expanding these hair follicular cells and advancing development of cell-based therapy for both wound healing and hair regeneration. The current study supports that optimized oxygen tension can be applied to use expanded human DPCs and DSCs for skin engineering and clinical applications.
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Affiliation(s)
- Koji Kanayama
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan.,Department of Plastic Surgery, The University of Tokyo School of Medicine, Tokyo, Japan
| | - Hitomi Takada
- Laboratory of Stem Cell Technology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma City, Japan
| | - Natsumi Saito
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Harunosuke Kato
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Kahori Kinoshita
- Department of Plastic Surgery, The University of Tokyo School of Medicine, Tokyo, Japan
| | - Takako Shirado
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Takanobu Mashiko
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan.,Department of Plastic Surgery, The University of Tokyo School of Medicine, Tokyo, Japan
| | - Rintaro Asahi
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Masanori Mori
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Kensuke Tashiro
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Ataru Sunaga
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
| | - Akira Kurisaki
- Laboratory of Stem Cell Technology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma City, Japan
| | - Katsutoshi Yoshizato
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan.,Synthetic Biology Laboratory, Graduate School of Medicine, Osaka City University, Osaka, Japan
| | - Kotaro Yoshimura
- Department of Plastic Surgery, Jichi Medical University, Shimotsuke City, Japan
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40
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Tissue Engineering and Regenerative Medicine in Craniofacial Reconstruction and Facial Aesthetics. J Craniofac Surg 2020; 31:15-27. [PMID: 31369496 DOI: 10.1097/scs.0000000000005840] [Citation(s) in RCA: 34] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
The craniofacial region is anatomically complex and is of critical functional and cosmetic importance, making reconstruction challenging. The limitations of current surgical options highlight the importance of developing new strategies to restore the form, function, and esthetics of missing or damaged soft tissue and skeletal tissue in the face and cranium. Regenerative medicine (RM) is an expanding field which combines the principles of tissue engineering (TE) and self-healing in the regeneration of cells, tissues, and organs, to restore their impaired function. RM offers many advantages over current treatments as tissue can be engineered for specific defects, using an unlimited supply of bioengineered resources, and does not require immunosuppression. In the craniofacial region, TE and RM are being increasingly used in preclinical and clinical studies to reconstruct bone, cartilage, soft tissue, nerves, and blood vessels. This review outlines the current progress that has been made toward the engineering of these tissues for craniofacial reconstruction and facial esthetics.
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41
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Zhang K, Bai X, Yuan Z, Cao X, Jiao X, Qin Y, Wen Y, Zhang X. Cellular Nanofiber Structure with Secretory Activity-Promoting Characteristics for Multicellular Spheroid Formation and Hair Follicle Regeneration. ACS APPLIED MATERIALS & INTERFACES 2020; 12:7931-7941. [PMID: 32003218 DOI: 10.1021/acsami.9b21125] [Citation(s) in RCA: 27] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/10/2023]
Abstract
Multicellular spheroids can mimic the in vivo microenvironment and maintain the unique functions of tissues, which has attracted great attention in tissue engineering. However, the traditional culture microenvironment with structural deficiencies complicates the culture and collection process and tends to lose the function of multicellular spheroids with the increase of cell passage. In order to construct efficient and functional multicellular spheroids, in this study, a chitosan/polyvinyl alcohol nanofiber sponge which has an open-cell cellular structure is obtained. The hair follicle (HF) regeneration model was employed to evaluate HF-inducing ability of dermal papilla (DP) multicellular spheroids which formed on the cellular structure nanofiber sponge. Through structural fine-tuning, the nanofiber sponge has appropriate elasticity for the creation of a three-dimensional dynamic microenvironment to regulate cellular behavior. The cellular structure nanofiber sponge tilts the balance of cell-substratum and cell-cell interactions to a state which is more conducive to the formation of controllable multicellular spheroids in a short time. More importantly, it improves the secretory activity of high-passaged dermal papilla cells and restores their intrinsic properties. Experiments using BALB/c nude mice show that cultured DP multicellular spheroids could effectively enhance HF-inducing ability. This novel system provides a simple and efficient strategy for multicellular spheroid formation and HF regeneration.
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Affiliation(s)
- Kexin Zhang
- Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering , University of Science and Technology Beijing , Beijing 100083 , P. R. China
| | - Xiufeng Bai
- Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics , Chinese Academy of Sciences , 15 Datun Road , Chaoyang District, Beijing 100101 , China
| | - Zhipeng Yuan
- Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering , University of Science and Technology Beijing , Beijing 100083 , P. R. China
| | - Xintao Cao
- Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics , Chinese Academy of Sciences , 15 Datun Road , Chaoyang District, Beijing 100101 , China
- University of Chinese Academy of Sciences , Beijing 100049 , China
| | - Xiangyu Jiao
- Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering , University of Science and Technology Beijing , Beijing 100083 , P. R. China
| | - Yan Qin
- Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics , Chinese Academy of Sciences , 15 Datun Road , Chaoyang District, Beijing 100101 , China
| | - Yongqiang Wen
- Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering , University of Science and Technology Beijing , Beijing 100083 , P. R. China
| | - Xueji Zhang
- Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering , University of Science and Technology Beijing , Beijing 100083 , P. R. China
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42
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Chen Y, Huang J, Chen R, Yang L, Wang J, Liu B, Du L, Yi Y, Jia J, Xu Y, Chen Q, Ngondi DG, Miao Y, Hu Z. Sustained release of dermal papilla-derived extracellular vesicles from injectable microgel promotes hair growth. Am J Cancer Res 2020; 10:1454-1478. [PMID: 31938074 PMCID: PMC6956798 DOI: 10.7150/thno.39566] [Citation(s) in RCA: 46] [Impact Index Per Article: 9.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/23/2019] [Accepted: 11/07/2019] [Indexed: 02/06/2023] Open
Abstract
Hair regeneration has long captured researchers' attention because alopecia is a common condition and current therapeutic approaches have significant limitations. Dermal papilla (DP) cells serve as a signaling center in hair follicles and regulate hair formation and cycling by paracrine secretion. Secreted EVs are important signaling mediators for intercellular communication, and DP-derived extracellular vesicles (DP-EVs) may play an important role in hair regeneration. However, the instability of EVs in vivo and their low long-term retention after transplantation hinder their use in clinical applications. Methods: Human DP-EVs were encapsulated in partially oxidized sodium alginate (OSA) hydrogels, yielding OSA-encapsulated EVs (OSA-EVs), which act as a sustained-release system to increase the potential therapeutic effect of DP-EVs. The ability of the OSA-EVs to protect protein was assessed. The hair regeneration capacity of OSA-EVs, as well as the underlying mechanism, was explored in hair organ culture and a mouse model of depilation. Results: The OSA-EVs were approximately 100 μm in diameter, and as the hydrogel degraded, DP-EVs were gradually released. In addition, the hydrogel markedly increased the stability of vesicular proteins and increased the retention of EVs in vitro and in vivo. The OSA-EVs significantly facilitated proliferation of hair matrix cells, prolonged anagen phase in cultured human hairs, and accelerated the regrowth of back hair in mice after depilation. These effects may be due to upregulation of hair growth-promoting signaling molecules such as Wnt3a and β-catenin, and downregulation of inhibitory molecule BMP2. Conclusion: This study demonstrated that OSA hydrogels promote the therapeutic effects of DP-EVs, and indicate that our novel OSA-EVs could be used to treat alopecia.
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43
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Su Y, Wen J, Zhu J, Xie Z, Liu C, Ma C, Zhang Q, Xu X, Wu X. Pre-aggregation of scalp progenitor dermal and epidermal stem cells activates the WNT pathway and promotes hair follicle formation in in vitro and in vivo systems. Stem Cell Res Ther 2019; 10:403. [PMID: 31856904 PMCID: PMC6921573 DOI: 10.1186/s13287-019-1504-6] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2019] [Revised: 11/12/2019] [Accepted: 11/20/2019] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Billions of dollars are invested annually by pharmaceutical companies in search of new options for treating hair loss conditions; nevertheless, the challenge remains. One major limitation to hair follicle research is the lack of effective and efficient drug screening systems using human cells. Organoids, three-dimensional in vitro structures derived from stem cells, provide new opportunities for studying organ development, tissue regeneration, and disease pathogenesis. The present study focuses on the formation of human hair follicle organoids. METHODS Scalp-derived dermal progenitor cells mixed with foreskin-derived epidermal stem cells at a 2:1 ratio aggregated in suspension to form hair follicle-like organoids, which were confirmed by immunostaining of hair follicle markers and by molecular dye labeling assays to analyze dermal and epidermal cell organization in those organoids. The hair-forming potential of organoids was examined using an in vivo transplantation assay. RESULTS Pre-aggregation of dermal and epidermal cells enhanced hair follicle formation in vivo. In vitro pre-aggregation initiated the interactions of epidermal and dermal progenitor cells resulting in activation of the WNT pathway and the formation of pear-shape structures, named type I aggregates. Cell-tracing analysis showed that the dermal and epidermal cells self-assembled into distinct epidermal and dermal compartments. Histologically, the type I aggregates expressed early hair follicle markers, suggesting the hair peg-like phase of hair follicle morphogenesis. The addition of recombinant WNT3a protein to the medium enhanced the formation of these aggregates, and the Wnt effect could be blocked by the WNT inhibitor, IWP2. CONCLUSIONS In summary, our system supports the rapid formation of a large number of hair follicle organoids (type I aggregates). This system provides a platform for studying epithelial-mesenchymal interactions, for assessing inductive hair stem cells and for screening compounds that support hair follicle regeneration.
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Affiliation(s)
- Yiqun Su
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
- Department of Implantology, School and Hospital of Stomatology, Shandong University, Jinan, China
| | - Jie Wen
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
| | - Junrong Zhu
- Women and Children's Hospital of Hubei Province, Wuhan, Hubei, China
| | - Zhiwei Xie
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
- Department of Stomatology, Shengli Oilfield Central Hospital, Dongying, Shandong, China
| | - Chang Liu
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
| | - Chuan Ma
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
| | - Qun Zhang
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China
| | - Xin Xu
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China.
- Department of Implantology, School and Hospital of Stomatology, Shandong University, Jinan, China.
- School of Stomatology, Shandong University, 44-1 Wenhua West Road, Jinan, 250014, Shandong, China.
| | - Xunwei Wu
- Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Shandong University, Jinan, China.
- Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China.
- School of Stomatology, Shandong University, 44-1 Wenhua West Road, Jinan, 250014, Shandong, China.
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44
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Kazi T, Niibe I, Nishikawa A, Matsuzaki T. Optimal stimulation toward the dermal papilla lineage can be promoted by combined use of osteogenic and adipogenic inducers. FEBS Open Bio 2019; 10:197-210. [PMID: 31730301 PMCID: PMC6996385 DOI: 10.1002/2211-5463.12763] [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: 09/22/2018] [Revised: 04/01/2019] [Accepted: 11/13/2019] [Indexed: 11/22/2022] Open
Abstract
Dermal papilla cells (DPCs) play crucial roles in hair regeneration, but they readily lose their hair‐forming ability during in vitro culture. Although the formation of spheroids partially restores the ability, shrinkage of the spheroids makes it difficult to maintain cellular viability. To address this problem, we stimulated DPCs with factors known to induce adipogenic and/or osteogenic differentiation, because DPCs share unique gene expression profiles with adipocytes and osteocytes. We isolated DPCs from versican (vcan)–GFP mice, in which GFP is expressed under the control of a vcan promoter, which is strongly active in DPCs of anagen hair follicles. GFP fluorescence was most intense when the spheroids were made from DPCs cultured in a half‐diluted combination of adipogenic and osteogenic media (CAO1/2), a Dulbecco’s modified Eagle’s medium‐based medium that contains 10% FBS, 275 nm dexamethasone, 2.5 mm β‐glycerol phosphate, 12.5 µg·mL−1 ascorbic acid, 0.125 µm isobutylmethylxanthine and 2.5 ng·mL−1 insulin. The dose of each additive used was less than the optimal dose for adipogenic or osteogenic differentiation, and shrinkage of the spheroids was avoided through the addition of fibroblast growth factor 2 and platelet‐derived growth factor‐AA to CAO1/2. In addition, the gene and protein expression of vcan, osteopontin, alkaline phosphatase and α‐smooth muscle actin in the spheroids were augmented to levels similar to those of the intact dermal papillae, which exhibited restored hair‐forming activity. In conclusion, a combination of certain adipogenic and osteogenic inducers, together with fibroblast growth factor 2 and platelet‐derived growth factor‐AA, can promote differentiation toward the DPC lineage.
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Affiliation(s)
- Taheruzzaman Kazi
- Bioscience and Biotechnology, The United Graduate School of Agricultural Sciences, Tottori University, Japan
| | - Ichitaro Niibe
- Department of Biological Science, Faculty of Life and Environment Science, Shimane University, Japan
| | - Akio Nishikawa
- Bioscience and Biotechnology, The United Graduate School of Agricultural Sciences, Tottori University, Japan.,Department of Biological Science, Faculty of Life and Environment Science, Shimane University, Japan
| | - Takashi Matsuzaki
- Bioscience and Biotechnology, The United Graduate School of Agricultural Sciences, Tottori University, Japan.,Department of Biological Science, Faculty of Life and Environment Science, Shimane University, Japan
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45
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Stratoulias V, Michon F. Tooth bioengineering from single cell suspensions. MethodsX 2019; 6:2429-2438. [PMID: 31720232 PMCID: PMC6838984 DOI: 10.1016/j.mex.2019.10.009] [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: 08/21/2019] [Accepted: 10/08/2019] [Indexed: 11/18/2022] Open
Abstract
Recent advances in bioengineering and biomaterials, along with knowledge deriving from the fields of developmental biology and stem cell research, have rendered feasible functional replacement of full organs. Here, we describe the methodology for bioengineering a tooth, starting from embryonic epithelial and mesenchymal single cell suspensions. In addition, we describe the subsequent steps of processing this minute structure for use in applications such as histological examination, immunofluorescence and in situ hybridisation. This methodology can be used for any minute structure that needs to be used in paraffin blocks. •Detailed methodology for reproducible and reliable results•Extra step to ensure single cell populations•Subsequent minute structure processing for histological analysis.
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Affiliation(s)
- Vassilis Stratoulias
- Institute of Biotechnology, Helsinki Institute of Life Science, Developmental Biology Program, University of Helsinki, 00790, Helsinki, Finland
- Corresponding author.
| | - Frederic Michon
- Institute of Biotechnology, Helsinki Institute of Life Science, Developmental Biology Program, University of Helsinki, 00790, Helsinki, Finland
- Institute for Neurosciences of Montpellier, INSERM UMR1051, University of Montpellier, 34295, Montpellier, France
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46
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Tan JJY, Common JE, Wu C, Ho PCL, Kang L. Keratinocytes maintain compartmentalization between dermal papilla and fibroblasts in 3D heterotypic tri-cultures. Cell Prolif 2019; 52:e12668. [PMID: 31379046 PMCID: PMC6797517 DOI: 10.1111/cpr.12668] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/24/2019] [Revised: 06/11/2019] [Accepted: 06/27/2019] [Indexed: 01/01/2023] Open
Abstract
OBJECTIVES Reproducing human hair follicles in vitro is often limited by various reasons such as the lack of a systematic approach to culture distinct hair follicle cell types to reproduce their spatial relationship. Here, we reproduce hair follicle-like constructs resembling the spatial orientation of different cells in vivo, to study the role of keratinocytes in maintaining cellular compartmentalization among hair follicle-related cells. MATERIALS AND METHODS Dermal papilla (DP) cells, HaCaT keratinocytes and human dermal fibroblast (HDF) cells were seeded sequentially into three-dimensional (3D) microwells fabricated from polyethylene glycol diacrylate hydrogels. Quantitative polymerase chain reaction was used to compare inductive gene expression of 3D and two-dimensional (2D) DP. DP and HaCaT cells were transfected with green fluorescent protein and red fluorescent protein lentivirus, respectively, to enable cell visualization using confocal microscopy. RESULTS The 3D DP cultures showed significantly enhanced expression of essential DP genes as compared 2D cultures. Core-shell configurations containing keratinocytes forming the outer shell and DP forming the core were observed. Migratory polarization was mediated by cell-cell interaction between the keratinocytes and HDF cells, while preserving the aggregated state of the DP cells. CONCLUSIONS Keratinocytes may play a role in maintaining compartmentalization between the DP and the surrounding HDF residing in the dermis, and therefore maintains the aggregative state of the DP cells, necessary for hair follicle development and function.
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Affiliation(s)
- Justin J. Y. Tan
- Department of PharmacyNational University of SingaporeSingaporeSingapore
| | | | - Chunyong Wu
- Department of Pharmaceutical AnalysisChina Pharmaceutical UniversityNanjingChina
| | - Paul C. L. Ho
- Department of PharmacyNational University of SingaporeSingaporeSingapore
| | - Lifeng Kang
- School of PharmacyUniversity of SydneySydneyNSWAustralia
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Zhang X, Xiao S, Liu B, Miao Y, Hu Z. Use of extracellular matrix hydrogel from human placenta to restore hair-inductive potential of dermal papilla cells. Regen Med 2019; 14:741-751. [PMID: 31368409 DOI: 10.2217/rme-2018-0112] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022] Open
Abstract
Aim: To explore the feasibility of human placenta extracellular matrix (HPECM) hydrogel in restoring the hair-inductive capacity of high-passaged (P8) dermal papilla cells (DPCs) for hair follicle regeneration. Materials & methods: HPECM hydrogel was prepared following decellularization and enzymatic solubilization treatment. DPCs isolated from human scalp were cultured in 2D and 3D environments. The hair-inductive ability of DPCs was assessed by quantitative RT-PCR, immunofluorescence staining, immunoblotting and patch assay. Results: DPCs (P8) formed spheres when cultured on the HPECM hydrogel. The expression levels of Versican, ALP, and β-catenin were restored in the DP spheres. HPECM hydrogel-cultured DP spheres co-grafted with newborn mouse epidermal cells regenerated new hair follicle. Conclusion: HPECM hydrogel successfully restores the hair-inductive capacity of high-passaged DPCs.
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Affiliation(s)
- Xinyu Zhang
- Department of Plastic, Cosmetic & Maxillofacial Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi 'an, ShanXi, PR China.,Department of Plastic Surgery, Nan Fang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Shune Xiao
- Department of Plastic Surgery, Nan Fang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Bingcheng Liu
- Department of Plastic Surgery, Nan Fang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Yong Miao
- Department of Plastic Surgery, Nan Fang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China
| | - Zhiqi Hu
- Department of Plastic Surgery, Nan Fang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China
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48
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Zhao Q, Li N, Zhang H, Lei X, Cao Y, Xia G, Duan E, Liu S. Chemically induced transformation of human dermal fibroblasts to hair‐inducing dermal papilla‐like cells. Cell Prolif 2019. [DOI: doi.org/10.1111/cpr.12652] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/03/2022] Open
Affiliation(s)
- Qian Zhao
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
- State Key Laboratory of Agrobiotechnology, College of Biological Sciences China Agricultural University Beijing China
| | - Na Li
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
- University of Chinese Academy of Sciences Beijing China
| | - Huishan Zhang
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
| | - Xiaohua Lei
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
- State Key Laboratory of Agrobiotechnology, College of Biological Sciences China Agricultural University Beijing China
| | - Yujing Cao
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
| | - Guoliang Xia
- State Key Laboratory of Agrobiotechnology, College of Biological Sciences China Agricultural University Beijing China
| | - Enkui Duan
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
| | - Shuang Liu
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology Chinese Academy of Sciences Beijing China
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49
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Zhao Q, Li N, Zhang H, Lei X, Cao Y, Xia G, Duan E, Liu S. Chemically induced transformation of human dermal fibroblasts to hair-inducing dermal papilla-like cells. Cell Prolif 2019; 52:e12652. [PMID: 31264301 PMCID: PMC6797507 DOI: 10.1111/cpr.12652] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/31/2019] [Revised: 05/09/2019] [Accepted: 05/13/2019] [Indexed: 12/25/2022] Open
Affiliation(s)
- Qian Zhao
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.,State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China
| | - Na Li
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.,University of Chinese Academy of Sciences, Beijing, China
| | - Huishan Zhang
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
| | - Xiaohua Lei
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.,State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China
| | - Yujing Cao
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
| | - Guoliang Xia
- State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China
| | - Enkui Duan
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
| | - Shuang Liu
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
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50
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Zheng M, Jang Y, Choi N, Kim DY, Han TW, Yeo JH, Lee J, Sung JH. Hypoxia improves hair inductivity of dermal papilla cells via nuclear NADPH oxidase 4-mediated reactive oxygen species generation'. Br J Dermatol 2019; 181:523-534. [PMID: 30703252 DOI: 10.1111/bjd.17706] [Citation(s) in RCA: 23] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/24/2019] [Indexed: 12/15/2022]
Abstract
BACKGROUND Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis. Therefore, tremendous efforts have been made to promote DPC hair inductivity. OBJECTIVES The aim of this study was to investigate the mitogenic and hair inductive effects of hypoxia on DPCs and examine the underlying mechanism of hypoxia-induced stimulation of DPCs. METHODS DPCs' hair inductivity was examined under normoxia (20% O2 ) and hypoxia (2% O2 ). RESULTS Hypoxia significantly increased the proliferation and delayed senescence of DPCs via Akt phosphorylation and downstream pathways. Hypoxia upregulated growth factor secretion of DPCs through the mitogen-activated protein kinase pathway. Hypoxia-preconditioned DPCs induced the telogen-to-anagen transition in C3 H mice, and also enhanced hair neogenesis in a hair reconstitution assay. Injected green fluorescent protein-labelled DPCs migrated to the outer root sheath of the hair follicle, and hypoxia-preconditioning increased survival and migration of DPCs in vivo. Conditioned medium obtained from hypoxia increased the hair length of mouse vibrissa follicles via upregulation of alkaline phosphatase, vascular endothelial growth factor, and glial cell line-derived neurotrophic factor. We examined the mechanism of this hypoxia-induced stimulation, and found that reactive oxygen species (ROS) play a key role. For example, inhibition of ROS generation by N-acetylcysteine or diphenyleneiodonium treatment attenuated DPCs' hypoxia-induced stimulation, but treatment with ROS donors induced mitogenic effects and anagen transition. NADPH oxidase 4 is highly expressed in the DPC nuclear region, and NOX4 knockout by CRISPR-Cas9 attenuated the hypoxia-induced stimulation of DPCs. CONCLUSIONS Our results suggest that DPC culture under hypoxia has great advantages over normoxia, and is a novel solution for producing DPCs for cell therapy. What's already known about this topic? Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis, but they are difficult to isolate and expand for use in cell therapy. Tremendous efforts have been made to increase proliferation of DPCs and promote their hair formation ability. What does this study add? Hypoxia (2% O2 ) culture of DPCs increases proliferation, delays senescence and enhances hair inductivity of DPCs. Reactive oxygen species play a key role in hypoxia-induced stimulation of DPC. What is the translational message? Preconditioning DPCs under hypoxia improves their hair regenerative potential, and is a novel solution for producing DPCs for cell therapy to treat hair loss.
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Affiliation(s)
- M Zheng
- STEMORE Co. Ltd, Incheon, South Korea
| | - Y Jang
- STEMORE Co. Ltd, Incheon, South Korea
| | - N Choi
- College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
| | - D Y Kim
- College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
| | - T W Han
- College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
| | - J H Yeo
- College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
| | - J Lee
- College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
| | - J-H Sung
- STEMORE Co. Ltd, Incheon, South Korea.,College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, South Korea
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