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Zhu D, Barabadi M, McDonald C, Kusuma G, Inocencio IM, Lim R. Implications of maternal-fetal health on perinatal stem cell banking. Gene Ther 2024; 31:65-73. [PMID: 37880336 PMCID: PMC10940157 DOI: 10.1038/s41434-023-00426-w] [Citation(s) in RCA: 1] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/13/2022] [Revised: 08/21/2023] [Accepted: 10/09/2023] [Indexed: 10/27/2023]
Abstract
Cell based therapies are being assessed for their therapeutic potential across a variety of diseases. Gestational tissues are attractive sources for cell therapy. The large number of births worldwide ensures sufficient access to gestational tissues, however, limited information has been reported around the impact of birth trends, delivery methods and pregnancy conditions on perinatal stem cell banking. This review describes the current state of banking of gestational tissues and their derived perinatal stem cells, discusses why the changes in birth trends and delivery methods could affect gestational tissue banking practices, and further explores how common pregnancy complications can potentially influence perinatal stem cell banking.
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Affiliation(s)
- Dandan Zhu
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia
| | - Mehri Barabadi
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia.
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia.
| | - Courtney McDonald
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia
| | - Gina Kusuma
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia
| | - Ishmael Miguel Inocencio
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia
| | - Rebecca Lim
- The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
- Department of Obstetrics and Gynaecology, Monash University, Monash, VIC, Australia
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Sharma R, Prakash S, Jain A, Pahwa D, Kalra J. Maternal and neonatal variables affecting CD34+ cell count in the umbilical cord blood. JOURNAL OF APPLIED HEMATOLOGY 2022. [DOI: 10.4103/joah.joah_68_21] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/04/2022] Open
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Di Tullio I, Azzolina D, Piras GN, Comoretto RI, Minto C, De Angeli S, Gregori D. Factors associated with blood cord unit bankability: an analysis of a 15-year-long case series. Cell Tissue Bank 2020; 21:77-87. [PMID: 31848776 DOI: 10.1007/s10561-019-09799-y] [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: 05/20/2019] [Accepted: 12/01/2019] [Indexed: 11/29/2022]
Abstract
Blood banking is a long and complex process requiring an accurate screening of potential donors and high-quality control systems. Previous studies in literature investigated factors potentially determining a higher cell levels with the aim of optimizing donors' selection and improving banking process. This study aims to identify factors associated with the concentration of stem cells in umbilical cord blood, so increasing the probability of bankability, focusing on the possible implications in terms of obstetric and resources management. This is a retrospective study conducted in the Obstetric Units of two Italian Hospitals in Montebelluna and Castelfranco Veneto. Study has been conducted on cord blood units banked between 1999 and 2015. Data on medical histories and clinical characteristics of mother and baby have been retrieved via a retrospective examination of medical records. A total of 869 cord blood units were studied. At multivariable analysis, in agreement with literature, birthweight and placental weight have been found to be associated with higher concentration of total nucleated cells. As additional factor, amount of fluid infused was associated with cord blood units' count. This study is the first one to clearly identify the role of fluid infusion on cord blood units' counts in addition to placental weight and delivery. Some non-modifiable features can help in predicting bankability from pre-natal aspects to factors more related with obstetric management is suggested.
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Affiliation(s)
- Isabella Di Tullio
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy
| | - Danila Azzolina
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy
| | - Gianluca Niccolò Piras
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy
| | - Rosanna Irene Comoretto
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy
| | - Clara Minto
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy
| | - Sergio De Angeli
- ULSS 9, Treviso, Viale Bartolomeo D'Alviano 34, 31100, Treviso, Italy
| | - Dario Gregori
- Unit of Biostatistics, Epidemiology and Public Health, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, University of Padova, Via Loredan 18, 35131, Padua, Italy.
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Mousavi SH, Zarrabi M, Abroun S, Ahmadipanah M, Abbaspanah B. Umbilical cord blood quality and quantity: Collection up to transplantation. Asian J Transfus Sci 2019; 13:79-89. [PMID: 31896912 PMCID: PMC6910041 DOI: 10.4103/ajts.ajts_124_18] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2018] [Accepted: 06/16/2019] [Indexed: 11/04/2022] Open
Abstract
Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells for transplantation in some blood disorders. One of the major factors that influence on transplantation fate is cord blood (CB) cell count, in addition to human leukocyte antigen similarity and CD34+ cell number. Here, we review the factors that could effect on quality and quantity of CBUs. Relevant English-language literatures were searched and retrieved from PubMed using the terms: CB, quality, collection, and transplantation. The numbers of total nucleated cells (TNCs) and CD34+ cells are good indicators of CB quality because they have been associated with engraftment; thereby, whatever the TNCs in a CB unit (CBU) are higher, more likely they led to successful engraftment. Many factors influence the quantity and quality of UCB units that collect after delivery. Some parameters are not in our hands, such as maternal and infant factors, and hence, we cannot change these. However, some other factors are in our authority, such as mode of collection, type and amount of anticoagulant, and time and temperature during collection to postthaw CBUs and freeze-and-thaw procedures. By optimizing the CB collection, we can improve the quantity and quality of UCB for storage and increase the likelihood of its use for transplantation.
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Affiliation(s)
- Seyed Hadi Mousavi
- Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
| | - Morteza Zarrabi
- Cord Blood Bank, Royan Stem Cell Technology Institute, Tehran, Iran
| | - Saeid Abroun
- Department of Hematology, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran
| | - Mona Ahmadipanah
- Cord Blood Bank, Royan Stem Cell Technology Institute, Tehran, Iran
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Maternal predictors and quality of umbilical cord blood units. Cell Tissue Bank 2017; 19:69-75. [PMID: 28823040 PMCID: PMC5829103 DOI: 10.1007/s10561-017-9657-y] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2017] [Accepted: 08/14/2017] [Indexed: 11/30/2022]
Abstract
The aim of the study was to determine the relationship between the maternal age at delivery and selected properties of the cord blood stem cells. The study included 50 pregnant women aged between 18 and 38 years in which spontaneous labors or elective cesarean sections were performed. Umbilical cord blood was collected immediately after the women were delivered of newborns. The samples were analyzed in the Polish Stem Cells Bank in Warsaw. The highest mean WBC level (p < 0.05) was observed in the umbilical blood collected from patients aged 35 years and more. Similarly, the highest mean cell viability was observed in the umbilical cord blood collected from patients aged 35 and more. There were no statistically significant correlations between the CD34+ cells count and mean cell viability in the umbilical cord blood and the maternal age. With the significance level at p < 0.001, the females after spontaneous labor revealed a visibly higher WBC level than patients after a cesarean section. The higher mean WBC concentration (24.95 thousand/μl) was observed in the umbilical cord blood of patients aged 35 and more after spontaneous labors. In the same group, the umbilical cord blood was also characterized by the highest mean cell viability (98.72%). The number of nucleated cells in the umbilical cord blood collected in the perinatal period increases together with the maternal age. In the course of physiological spontaneous labors, the collected umbilical cord blood has more nucleated cells as compared with elective caesarian sections.
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Mousavi SH, Abroun S, Zarrabi M, Ahmadipanah M. The effect of maternal and infant factors on cord blood yield. Pediatr Blood Cancer 2017; 64:e26381. [PMID: 27905684 DOI: 10.1002/pbc.26381] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/05/2016] [Revised: 11/05/2016] [Accepted: 11/07/2016] [Indexed: 11/11/2022]
Abstract
Umbilical cord blood (CB) can be used as an alternative hematopoietic stem cell source for transplantation in hematological malignancy and blood disorders. The success of transplantation is highly related to the levels of total nucleated cell and CD34+ cell counts. The evaluation of optimal conditions can decrease the rate of graft rejection due to low cell count and increases the quality of CB units (CBUs) in the blood bank and the success rate of engraftment. To this end, we review the maternal and infant parameters affecting the quality and quantity of CBUs.
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Affiliation(s)
- Seyed Hadi Mousavi
- Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
| | - Saeid Abroun
- Department of Hematology, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran
| | - Morteza Zarrabi
- Royan Stem Cell Technology Institute, Cord Blood Bank, Tehran, Iran
| | - Mona Ahmadipanah
- Royan Stem Cell Technology Institute, Cord Blood Bank, Tehran, Iran
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Habibollah S, Forraz N, McGuckin CP. Application of Umbilical Cord and Cord Blood as Alternative Modes for Liver Therapy. Regen Med 2015. [DOI: 10.1007/978-1-4471-6542-2_22] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023] Open
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Wisgrill L, Schüller S, Bammer M, Berger A, Pollak A, Radke TF, Kögler G, Spittler A, Helmer H, Husslein P, Gortner L. Hematopoietic stem cells in neonates: any differences between very preterm and term neonates? PLoS One 2014; 9:e106717. [PMID: 25181353 PMCID: PMC4152327 DOI: 10.1371/journal.pone.0106717] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2014] [Accepted: 08/07/2014] [Indexed: 01/14/2023] Open
Abstract
Background In the last decades, human full-term cord blood was extensively investigated as a potential source of hematopoietic stem and progenitor cells (HSPCs). Despite the growing interest of regenerative therapies in preterm neonates, only little is known about the biological function of HSPCs from early preterm neonates under different perinatal conditions. Therefore, we investigated the concentration, the clonogenic capacity and the influence of obstetric/perinatal complications and maternal history on HSPC subsets in preterm and term cord blood. Methods CD34+ HSPC subsets in UCB of 30 preterm and 30 term infants were evaluated by flow cytometry. Clonogenic assays suitable for detection of the proliferative potential of HSPCs were conducted. Furthermore, we analyzed the clonogenic potential of isolated HSPCs according to the stem cell marker CD133 and aldehyde dehydrogenase (ALDH) activity. Results Preterm cord blood contained a significantly higher concentration of circulating CD34+ HSPCs, especially primitive progenitors, than term cord blood. The clonogenic capacity of HSPCs was enhanced in preterm cord blood. Using univariate analysis, the number and clonogenic potential of circulating UCB HSPCs was influenced by gestational age, birth weight and maternal age. Multivariate analysis showed that main factors that significantly influenced the HSPC count were maternal age, gestational age and white blood cell count. Further, only gestational age significantly influenced the clonogenic potential of UCB HSPCs. Finally, isolated CD34+/CD133+, CD34+/CD133– and ALDHhigh HSPC obtained from preterm cord blood showed a significantly higher clonogenic potential compared to term cord blood. Conclusion We demonstrate that preterm cord blood exhibits a higher HSPC concentration and increased clonogenic capacity compared to term neonates. These data may imply an emerging use of HSPCs in autologous stem cell therapy in preterm neonates.
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Affiliation(s)
- Lukas Wisgrill
- Dept. of Pediatrics and Adolescent Medicine, Division of Neonatology, Paediatric Intensive Care & Neuropaediatrics, Medical University of Vienna, Vienna, Austria
| | - Simone Schüller
- Dept. of Pediatrics and Adolescent Medicine, Division of Neonatology, Paediatric Intensive Care & Neuropaediatrics, Medical University of Vienna, Vienna, Austria
| | - Markus Bammer
- Dept. of Pediatrics and Adolescent Medicine, Division of Neonatology, Paediatric Intensive Care & Neuropaediatrics, Medical University of Vienna, Vienna, Austria
| | - Angelika Berger
- Dept. of Pediatrics and Adolescent Medicine, Division of Neonatology, Paediatric Intensive Care & Neuropaediatrics, Medical University of Vienna, Vienna, Austria
| | - Arnold Pollak
- Dept. of Pediatrics and Adolescent Medicine, Division of Neonatology, Paediatric Intensive Care & Neuropaediatrics, Medical University of Vienna, Vienna, Austria
| | - Teja Falk Radke
- Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Medical Center, Duesseldorf, Germany
| | - Gesine Kögler
- Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Medical Center, Duesseldorf, Germany
| | - Andreas Spittler
- Department of Surgery, Research Labs & Core Facility Flow Cytometry, Medical University of Vienna, Vienna, Austria
| | - Hanns Helmer
- Dept. of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria
| | - Peter Husslein
- Dept. of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria
| | - Ludwig Gortner
- Dept. of Pediatrics and Neonatology, Saarland University, Homburg, Saar, Germany
- * E-mail:
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Basford C, Forraz N, Habibollah S, Hanger K, McGuckin C. The cord blood separation league table: a comparison of the major clinical grade harvesting techniques for cord blood stem cells. Int J Stem Cells 2014; 3:32-45. [PMID: 24855539 DOI: 10.15283/ijsc.2010.3.1.32] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 03/27/2010] [Indexed: 11/09/2022] Open
Abstract
BACKGROUND AND OBJECTIVES Well over 1 million Umbilical Cord Blood units (UCB) have been stored globally in the last 10 years. Already, over 20,000 transplants been performed using UCB for haematopoietic reconstitution alone, now this potential is joined by regenerative medicine. However, more needs to be known about processing of this stem cell source for it to reach full potential. METHODS AND RESULTS IN THIS STUDY WE EVALUATED FIVE SEPARATION METHODS: plasma depletion, density gradient, Hetastarch, a novel method known as PrepaCyte-CB and an automated centrifugal machine. Sepax gives the highest recovery of nucleated cells, an average of 78.8% (SD±21.36). When looking at CD34+ haematopoietic stem cells PrepaCyte-CB provided the greatest recovery at 74.47% (SD±8.89). For volume reduction density gradient was the most effective leaving 0.03×10(6) RBC/ml, 8 times more efficient than its nearest competitor PrepaCyte-CB (p<0.05). Finally PrepaCyte-CB processing left samples with the highest clonogenic potential after processing and more significantly after cryopreservation: 9.23 CFU/10(8) cells (SD±2.33), 1.5 fold more effective than its nearest rival Sepax (p<0.05). CONCLUSIONS PrepaCyte-CB was the most flexible method; the only processing type unaffected by volume. Results indicate that processing choice is important depending on your final intended use.
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Affiliation(s)
- Christina Basford
- Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, NE1 3BZ, United Kingdom
| | - Nicolas Forraz
- CTI-LYON, Cell Therapy Research Institute, Parc Technologique de Lyon-St Priest, Cèdre 1, 97 Allée Alexandre Borodine, 69800 SAINT PRIEST LYON, France
| | - Saba Habibollah
- Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, NE1 3BZ, United Kingdom
| | - Kendal Hanger
- Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, NE1 3BZ, United Kingdom
| | - Colin McGuckin
- CTI-LYON, Cell Therapy Research Institute, Parc Technologique de Lyon-St Priest, Cèdre 1, 97 Allée Alexandre Borodine, 69800 SAINT PRIEST LYON, France
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Pope B, Hokin B, Grant R. Effect of maternal iron status on the number of CD34+ stem cells harvested from umbilical cord blood. Transfusion 2014; 54:1876-80. [DOI: 10.1111/trf.12547] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2013] [Revised: 11/18/2013] [Accepted: 11/25/2013] [Indexed: 11/28/2022]
Affiliation(s)
- Belinda Pope
- Pathology Department; Sydney Adventist Hospital; Wahroonga NSW Australia
- Australasian Research Institute; Sydney Adventist Hospital; Wahroonga NSW Australia
- Faculty of Medicine; University of New South Wales; Kensington NSW Australia
| | - Bevan Hokin
- Pathology Department; Sydney Adventist Hospital; Wahroonga NSW Australia
- Sydney Medical School; University of Sydney; Camperdown NSW Australia
| | - Ross Grant
- Australasian Research Institute; Sydney Adventist Hospital; Wahroonga NSW Australia
- Faculty of Medicine; University of New South Wales; Kensington NSW Australia
- Sydney Medical School; University of Sydney; Camperdown NSW Australia
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Cord blood serum-based eye drops: the impact of donor haematological and obstetric factors on the variability of epidermal growth factor levels. BLOOD TRANSFUSION = TRASFUSIONE DEL SANGUE 2013; 12 Suppl 1:s44-50. [PMID: 24120597 DOI: 10.2450/2013.0115-13] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Subscribe] [Scholar Register] [Received: 04/05/2013] [Accepted: 06/12/2013] [Indexed: 01/17/2023]
Abstract
BACKGROUND Cord blood serum (CBS)-based eye drops are successfully used in corneal epithelial wound healing and are prepared to supply a known amount of epidermal growth factor (EGF). Product standardisation includes expensive EGF dosage in all cord blood (CB) units. The influence of donor obstetric and haematological characteristics on EGF content was evaluated, to exclude unsuitable CBS and pre-select those CB units able to provide the correct EGF supply for healing corneal wounds. MATERIALS AND METHODS Data were retrospectively collected from 135 donors included in the Emilia Romagna Cord Blood Bank records. Obstetric characteristics, parity and gestational age of the mother, sex, birth weight and Apgar score of the neonate, placental weight, duration of labour and mode of delivery were considered. Haematological characteristics, CD34+ cell number, and total nucleated cell, white blood cell and platelet counts were recorded. EGF content in CB units was estimated by enzyme-linked immunosorbent assay. Statistical evaluation was performed by Mann-Whitney unpaired and Student's t tests. Correlations between variables were evaluated by using Pearson's (r) or Spearman's (ρ) correlation coefficients. RESULTS EGF content was significantly higher in CBS from donors aged <30 years and after vaginal deliveries as compared with scheduled Caesarean sections (1,386±580 vs 1,106±391 pg/mL; P=0.002). EGF content was significantly correlated with duration of labour (r=0.45; P=0.0001), number of CD34+ cells/mL (r=0.3; P=0.002) particularly in vaginal deliveries (r=0.36; P=0.003), mother's age (-0.25; P=0.005), neonate's birth weight (r=0.27; P=0.005), and total nucleated cell (r=0.25; P=0.006), white cell (r=0.29; P=0.001) and platelet (r=0.24; P=0.009) counts. No significant correlations were found between EGF content and parity, gestational age, placental weight, neonate's sex or Apgar scores. DISCUSSION EGF levels are higher in CB units from younger mothers (<30 years), with longer labour duration (>6 hours), and higher CD34+ cell content (>0.05×10(6)/mL). In order to optimise the preparation and costs of CBS-based eye drops, pre-selection of CB units is recommended.
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Insights and hopes in umbilical cord blood stem cell transplantations. J Biomed Biotechnol 2012; 2012:572821. [PMID: 23258957 PMCID: PMC3509718 DOI: 10.1155/2012/572821] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/05/2012] [Accepted: 10/08/2012] [Indexed: 11/17/2022] Open
Abstract
Over 20.000 umblical cord blood transplantations (UCBT) have been carried out around the world. Indeed, UCBT represents an attractive source of donor hematopoietic stem cells (HSCs) and, offer interesting features (e.g., lower graft-versus-host disease) compared to bone marrow transplantation (BMT). Thereby, UCBT often represents the unique curative option against several blood diseases. Recent advances in the field of UCBT, consisted to develop strategies to expand umbilical stem cells and shorter the timing of their engraftment, subsequently enhancing their availability for enhanced efficacy of transplantation into indicated patients with malignant diseases (e.g., leukemia) or non-malignant diseases (e.g., thalassemia major). Several studies showed that the expansion and homing of UCBSCs depends on specific biological factors and cell types (e.g., cytokines, neuropeptides, co-culture with stromal cells). In this review, we extensively present the advantages and disadvantages of current hematopoietic stem cell transplantations (HSCTs), compared to UBCT. We further describe the importance of cord blood content and obstetric factors on cord blood selection, and report the recent approaches that can be undertook to improve cord blood stem cell expansion as well as engraftment. Eventually, we provide two majors examples underlining the importance of UCBT as a potential cure for blood diseases.
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Paspala SA, Vishwakarma SK, Murthy TV, Rao TN, Khan AA. Potential role of stem cells in severe spinal cord injury: current perspectives and clinical data. STEM CELLS AND CLONING-ADVANCES AND APPLICATIONS 2012; 5:15-27. [PMID: 24198535 PMCID: PMC3781762 DOI: 10.2147/sccaa.s28477] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Stem cell transplantation for spinal cord injury (SCI) along with new pharmacotherapy research offers the potential to restore function and ease the associated social and economic burden in the years ahead. Various sources of stem cells have been used in the treatment of SCI, but the most convincing results have been obtained with neural progenitor cells in preclinical models. Although the use of cell-based transplantation strategies for the repair of chronic SCI remains the long sought after holy grail, these approaches have been to date the most successful when applied in the subacute phase of injury. Application of cell-based strategies for the repair and regeneration of the chronically injured spinal cord will require a combinational strategy that may need to include approaches to overcome the effects of the glial scar, inhibitory molecules, and use of tissue engineering strategies to bridge the lesion. Nonetheless, cell transplantation strategies are promising, and it is anticipated that the Phase I clinical trials of some form of neural stem cell-based approach in SCI will commence very soon.
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Affiliation(s)
- Syed Ab Paspala
- PAN Research Foundation, CARE, Hyderabad, India ; The Institute of Medical Sciences, Hyderabad, India
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Tschiedel S, Bach E, Jilo A, Wang SY, Lange T, Al-Ali HK, Vucinic V, Niederwieser D, Cross M. Bcr–Abl dependent post-transcriptional activation of NME2 expression is a specific and common feature of chronic myeloid leukemia. Leuk Lymphoma 2012; 53:1569-76. [DOI: 10.3109/10428194.2012.656631] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
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Meyer-Monard S, Tichelli A, Troeger C, Arber C, de Faveri GN, Gratwohl A, Roosnek E, Surbek D, Chalandon Y, Irion O, Castelli D, Passweg J, Kindler V. Initial cord blood unit volume affects mononuclear cell and CD34+ cell-processing efficiency in a non-linear fashion. Cytotherapy 2012; 14:215-22. [DOI: 10.3109/14653249.2011.634404] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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Associations among birth weight, placental weight, gestational period and product quality indicators of umbilical cord blood units. Transfus Apher Sci 2011; 46:39-45. [PMID: 22206793 DOI: 10.1016/j.transci.2011.10.031] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2011] [Revised: 08/17/2011] [Accepted: 10/19/2011] [Indexed: 11/21/2022]
Abstract
INTRODUCTION Numbers of CD34+ cell and total nucleated cell (TNC) and cord blood volume are commonly used as indicators for haematopoietic potential of umbilical cord blood (UCB) units. The purpose of this study was to investigate the relationship between donor-related factors and the quality indicators of UCB. METHODS Obstetric and neonatal clinical laboratory data of a total of 1549 UCB units were obtained from Buddhist Tzu Chi Stem Cells Center (BTCSCC) Cord Blood Bank. A retrospective multivariate analysis was conducted to analyze the data. RESULTS Our results showed that birth weight had positive correlations with each of the clinical features of CD34+ cell number, TNC count and unit volume of UCB, followed by the placental weight. Longer gestational period would decrease CD34+ cell number and volume of UCB. Female baby and mode of vaginal delivery of neonates were found to have larger amount of TNC in UCB. CONCLUSION Our results would be helpful and beneficial in building up standard criteria for evaluating stored UCB units.
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Azouna NB, Berraeis L, Regaya Z, Jenhani F. Immunophenotyping of hematopoietic progenitor cells: Comparison between cord blood and adult mobilized blood grafts. World J Stem Cells 2011; 3:104-12. [PMID: 22180831 PMCID: PMC3240678 DOI: 10.4252/wjsc.v3.i11.104] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/30/2011] [Revised: 09/30/2011] [Accepted: 10/05/2011] [Indexed: 02/06/2023] Open
Abstract
AIM To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis. METHODS First, we performed a qualitative analysis of CD34+ cell sub-populations in both CB and PAPB grafts using the standardized ISHAGE protocol and a wide panel of 20 monoclonal antibodies. Next, we studied some parameters, such as the age of mothers and the weight of newborns, which can influence the quality and the quantity of CD34+ cells from CB. RESULTS We found that the percentage of apoptotic cells was high in CB in comparison to PAPB (PAPB: 4.6% ± 2.6% vs CB: 53.4% ± 5.2%, P < 0.001). In CB, the weight of newborn and the age of the mother have the influence on CD34+ cells. The follow-up of Ag CD133 in the ISHAGE double platform protocol in association with CD45, CD34 and the 7'AAD shows an equal rate between the two cell populations CD133+CD45+CD34+ high and CD34+CD45+ high with a higher percentage. So, is the inclusion of Ac CD133 necessary in the present panel included in the ISHAGE method? Last part, we showed a significant presence of interferon γ in CB in comparison to PAPB, the annexin showing the high number of apoptotic cells in CB. CONCLUSION This study demonstrates that many different obstetric factors must be taken into account when processing and cryo-banking umbilical CB units for transplantation.
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Affiliation(s)
- Nesrine Ben Azouna
- Nesrine Ben Azouna, Faouzi Jenhani, Immunology Research Unit, Faculty of Pharmacy of Monastir, Monsatir University, TU-5000 Monastir, Tunisia
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Horváthová M, Jahnová E, Palkovičová L, Trnovec T, Hertz-Picciotto I. Dynamics of lymphocyte subsets in children living in an area polluted by polychlorinated biphenyls. J Immunotoxicol 2011; 8:333-45. [PMID: 22013978 DOI: 10.3109/1547691x.2011.615767] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Immune system development, particularly in the pre-natal and early post-natal periods, has far-reaching health consequences during childhood, as well as throughout life. Exposure to poly-chlorinated biphenyls (PCBs) during pre-natal and early life has been previously associated with changes in the incidence of infectious and allergic diseases in children, and humoral immunity alterations. Lymphocyte immunophenotyping is an important tool in the diagnosis of immunologic and hematologic disorders. This study used a lysed whole blood method for analysis of lymphocyte sub-populations in samples from children born and living in two districts: a highly-contaminated area (Michalovce) and one (Svidnik/Stropkov) with ≈ 2-fold lower environmental PCB levels. The percentages of B-lymphocytes (CD19(+)), activated HLADR(+)CD19(+) cells, and CD8(+) T-lymphocytes significantly increased at 6- and 16-months-of-age in both selected regions as compared to in cord blood values (p < 0.001). Levels of CD3(+) cells increased significantly (from 61 to 65%) in samples from Michalovce (p < 0.01). Levels of CD4(+) T-lymphocytes declined 10% among 16-month-olds in both regions (Michalovce at p < 0.001 and Svidnik/Stropkov at p < 0.01). Natural killer (NK) cell levels decreased 50% in Michalovce 6- and 16-month-old children and 42% among 6-month-olds in Svidnik/Stropkov (p < 0.001). Compared with the less-contaminated region, Michalovce samples showed significantly higher expression of CD3(+) T-lymphocytes, B-lymphocytes, and activated B-lymphocytes, whereas NK cells were less expressed. Even after adjustment for selected covariates, e.g., maternal cigarette smoking, age, parity, ethnicity, birth weight, and gender of infant, the levels of CD19(+), HLADR(+)CD19(+), and CD3(-)CD(16 + 56)(+) cells were seen to remain significantly different between the districts. These results showed that early-life environmental PCB exposure was associated with fluctuations in major lymphocyte subsets in children, suggesting that there is a post-natal immune system response to PCB exposures.
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Affiliation(s)
- M Horváthová
- Department of Immunology and Immunotoxicology, Bratislava, Slovakia.
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Slovinska L, Novotna I, Kubes M, Radonak J, Jergova S, Cigankova V, Rosocha J, Cizkova D. Umbilical Cord Blood Cells CD133+/CD133− Cultivation in Neural Proliferation Media Differentiates Towards Neural Cell Lineages. Arch Med Res 2011; 42:555-62. [DOI: 10.1016/j.arcmed.2011.10.003] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/07/2011] [Accepted: 09/26/2011] [Indexed: 01/10/2023]
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Coldwell KE, Lee SJ, Kean J, Khoo CP, Tsaknakis G, Smythe J, Watt SM. Effects of obstetric factors and storage temperatures on the yield of endothelial colony forming cells from umbilical cord blood. Angiogenesis 2011; 14:381-92. [PMID: 21720855 PMCID: PMC3155043 DOI: 10.1007/s10456-011-9222-4] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/03/2011] [Accepted: 06/16/2011] [Indexed: 11/08/2022]
Abstract
As umbilical cord blood (UCB) is a rich source of endothelial colony-forming cells (ECFC), our aim was twofold: (1) to examine potential obstetric selection criteria for achieving the highest ECFC yields from UCB units, and (2) to determine whether transient storage temperatures of fresh UCB and cryopreservation of UCB units affected ECFC yield and function. ECFC quality was assessed before and after cryopreservation by their clonogenic proliferative potential. Of the 20 factors examined, placental weight was the only statistically significant obstetric factor that predicted ECFC frequency in UCB. Studies on the effects of storage revealed that transient storage of fresh UCB at 4°C reduced ECFC yield compared with storage at 22°C, while cryopreservation of UCB MNCs significantly reduced ECFC recoveries. To our knowledge, this is the first demonstration that placental weight and temperature of storage prior to processing or culture have significant effects on ECFC frequency in UCB. Our studies further support the evidence that cryopreservation of UCB MNCs compromises ECFC recovery.
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Affiliation(s)
- Kate E. Coldwell
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
| | - Stephanie J. Lee
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
- Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, OX3 9DU UK
- Medical Sciences Division, University of Melbourne, Melbourne, 3050 Australia
| | - Jennifer Kean
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
- Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, OX3 9DU UK
- Department of Plastic and Reconstructive Surgery, Stoke Mandeville Hospital, Aylesbury, HP21 8AL UK
| | - Cheen P. Khoo
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
| | | | - Jon Smythe
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
| | - Suzanne M. Watt
- Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
- Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, OX3 9DU UK
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Lee HR, Park JS, Shin S, Roh EY, Yoon JH, Han KS, Kim BJ, Storms RW, Chao NJ. Increased numbers of total nucleated and CD34+ cells in blood group O cord blood: an analysis of neonatal innate factors in the Korean population. Transfusion 2011; 52:76-81. [PMID: 21790633 DOI: 10.1111/j.1537-2995.2011.03262.x] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
BACKGROUND We analyzed neonatal factors that could affect hematopoietic variables of cord blood (CB) donated from Korean neonates. STUDY DESIGN AND METHODS The numbers of total nucleated cells (TNCs), CD34+ cells, and CD34+ cells/TNCs of CB in neonates were compared according to sex, gestational age, birth weight, birth weight centile for gestational age, and ABO blood group. RESULTS With 11,098 CB units analyzed, blood group O CB showed an increased number of TNCs, CD34+ cells, and CD34+ cells/TNCs compared with other blood groups. Although TNC counts were lower in males, no difference in the number of CD34+ cells was demonstrated because the number of CD34+ cells/TNCs was higher in males. An increase in the gestational age resulted in an increase in the number of TNCs and decreases in the number of CD34+ cells and CD34+ cells/TNCs. The numbers of TNCs, CD34+ cells, and CD34+ cells/TNCs increased according to increased birth weight centile as well as birth weight. CONCLUSION CB with blood group O has unique hematologic variables in this large-scale analysis of Korean neonates, although the impact on the storage policies of CB banks or the clinical outcome of transplantation remains to be determined.
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Affiliation(s)
- Hye Ryun Lee
- Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea
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Forraz N, McGuckin CP. The umbilical cord: a rich and ethical stem cell source to advance regenerative medicine. Cell Prolif 2011; 44 Suppl 1:60-9. [PMID: 21481046 PMCID: PMC6495455 DOI: 10.1111/j.1365-2184.2010.00729.x] [Citation(s) in RCA: 66] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2010] [Accepted: 08/11/2010] [Indexed: 02/04/2023] Open
Abstract
Science and medicine place a lot of hope in the development of stem cell research and regenerative medicine. This review will define the concept of regenerative medicine and focus on an abundant stem cell source - neonatal tissues such as the umbilical cord. Umbilical cord blood has been used clinically for over 20 years as a cell source for haematopoietic stem cell transplantation. Beyond this, cord blood and umbilical cord-derived stem cells have demonstrated potential for pluripotent lineage differentiation (liver, pancreatic, neural tissues and more) in vitro and in vivo. This promising research has opened up a new era for utilization of neonatal stem cells, now used beyond haematology in clinical trials for autoimmune disorders, cerebral palsy or type I diabetes.
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Affiliation(s)
- N. Forraz
- CTI‐LYON, Cell Therapy Research Institute, Parc Technologique de Lyon St Priest, St Priest‐Lyon, France
| | - C. P. McGuckin
- CTI‐LYON, Cell Therapy Research Institute, Parc Technologique de Lyon St Priest, St Priest‐Lyon, France
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Chirumbolo S, Ortolani R, Veneri D, Raffaelli R, Peroni D, Pigozzi R, Colombatti M, Vella A. Lymphocyte phenotypic subsets in umbilical cord blood compared to peripheral blood from related mothers. CYTOMETRY PART B-CLINICAL CYTOMETRY 2011; 80:248-53. [PMID: 21692178 DOI: 10.1002/cyto.b.20588] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/26/2010] [Revised: 12/27/2010] [Accepted: 01/06/2011] [Indexed: 11/07/2022]
Affiliation(s)
- Salvatore Chirumbolo
- Department of Pathology and Diagnostics, Section of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy.
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Song K, Yin Y, Lv C, Liu T, Macedo HM, Fang M, Shi F, Ma X, Cui Z. Co-culture of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood using human autoserum. ASIA-PAC J CHEM ENG 2010. [DOI: 10.1002/apj.507] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022]
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Basford C, Forraz N, McGuckin C. Optimized multiparametric immunophenotyping of umbilical cord blood cells by flow cytometry. Nat Protoc 2010; 5:1337-46. [PMID: 20595961 DOI: 10.1038/nprot.2010.88] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
Umbilical cord blood is a key source of stem cells for transplantation and regenerative medicine. To maximize each cord blood sample, it is important to analyze its cellular populations. Many cord blood banks focus on counts of total nucleated cells and/or cells that carry the CD34 antigen, but this limited focus does not give a true estimation of cord blood content, quality and appropriateness for use. This protocol is the first of its kind to enable a comprehensive investigation of cord blood cellular populations. Using multicolor flow cytometry it is possible to examine expression of 26 antigens--including hematopoietic markers CD45 and CD34, immune markers CD19 and CD3, and HLA--using a total of only 1 x 10(6) cells from each unit for both pre- and post-processing. The samples are stained, lysed, washed and analyzed flow cytometrically. This method also provides valuable information beyond cord blood composition and quality; for example, it can also be used to assess whether maternal factors affect CD34(+) cell numbers. Collection, processing, cryopreservation and initial flow cytometry take approximately 5 h.
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Affiliation(s)
- Christina Basford
- Stem Cell Group, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, United Kingdom
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Hassanein SMA, Amer HA, Shehab AA, Hellal MMKH. Umbilical cord blood CD45+/CD34+cells coexpression in preterm and full-term neonates: a pilot study. J Matern Fetal Neonatal Med 2010; 24:229-33. [DOI: 10.3109/14767058.2010.482606] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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Abstract
OBJECTIVE To investigate the cost-effectiveness of private umbilical cord blood banking. METHODS A decision-analytic model was designed comparing private umbilical cord blood banking with no umbilical cord blood banking. Baseline assumptions included a cost of $3,620 for umbilical cord blood banking and storage for 20 years, a 0.04% chance of requiring an autologous stem cell transplant, a 0.07% chance of a sibling requiring an allogenic stem cell transplant, and a 50% reduction in risk of graft-versus-host disease if a sibling uses banked umbilical cord blood. RESULTS Private cord blood banking is not cost-effective because it cost an additional $1,374,246 per life-year gained. In sensitivity analysis, if the cost of umbilical cord blood banking is less than $262 or the likelihood of a child needing a stem cell transplant is greater than 1 in 110, private umbilical cord blood banking becomes cost-effective. CONCLUSION Currently, private umbilical cord blood banking is cost-effective only for children with a very high likelihood of needing a stem cell transplant. Patients considering private blood banking should be informed of the remote likelihood that a unit will be used for a child or another family member. LEVEL OF EVIDENCE III.
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Fievet N, Varani S, Ibitokou S, Briand V, Louis S, Perrin RX, Massougbogji A, Hosmalin A, Troye-Blomberg M, Deloron P. Plasmodium falciparum exposure in utero, maternal age and parity influence the innate activation of foetal antigen presenting cells. Malar J 2009; 8:251. [PMID: 19889240 PMCID: PMC2780449 DOI: 10.1186/1475-2875-8-251] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2009] [Accepted: 11/05/2009] [Indexed: 12/14/2022] Open
Abstract
Background Malaria in pregnancy is associated with immunological abnormalities in the newborns, such as hampered T-helper 1 responses and increased T-regulatory responses, while the effect of maternal Plasmodium falciparum infection on foetal innate immunity is still controversial. Materials and methods The immunophenotype and cytokine release by dendritic cells (DC) and monocytes were evaluated in cord blood from 59 Beninese women with or without malaria infection by using flow cytometry. Results Accumulation of malaria pigment in placenta was associated with a partial maturation of cord blood myeloid and plasmacytoid DC, as reflected by an up-regulated expression of the major histocompatibility complex class II molecules, but not CD86 molecules. Cells of newborns of mothers with malaria pigment in their placenta also exhibited significantly increased cytokine responses upon TLR9 stimulation. In addition, maternal age and parity influenced the absolute numbers and activation status of cord blood antigen-presenting cells. Lastly, maternal age, but not parity, influenced TLR3, 4 and 9 responses in cord blood cells. Discussion Our findings support the view that placental parasitization, as indicated by the presence of malaria pigment in placental leukocytes, is significantly associated with partial maturation of different DC subsets and also to slightly increased responses to TLR9 ligand in cord blood. Additionally, other factors, such as maternal age and parity should be taken into consideration when analysing foetal/neonatal innate immune responses. Conclusion These data advocate a possible mechanism by which PAM may modulate foetal/neonatal innate immunity.
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Affiliation(s)
- Nadine Fievet
- UR010, Mother and Child Health in the Tropics, Institut de Recherche pour le Développement (IRD), Cotonou, Benin
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Basford C, Forraz N, Habibollah S, Hanger K, McGuckin CP. Umbilical cord blood processing using Prepacyte-CB increases haematopoietic progenitor cell availability over conventional Hetastarch separation. Cell Prolif 2009; 42:751-61. [PMID: 19758367 DOI: 10.1111/j.1365-2184.2009.00646.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022] Open
Abstract
BACKGROUND Currently the most frequently used method for umbilical cord blood separation in many hospitals across the UK and the rest of the world, where small-to-medium amounts of samples are processed, is Hetastarch, a mechanical, starch-based method, which causes red cell agglutination by rouleaux formation. AIM In this study, a novel method (Prepa-Cyte-CB), in comparison with Hetastarch as part of an FDA-approved clinical study, was evaluated. MATERIALS AND METHODS Validation of data included recovery of nucleated and CD34+ cells, red blood cell reduction, colony forming unit potential, flow cytometric analysis and sterility tests. RESULTS PrepaCyte-CB, in comparison with Hetastarch offers fast, reliable separation with improved recovery of nucleated cells, 72.03% (+/-8.48 SD) compared to 58.09% (+/-20.06 SD), and CD34+ haematopoietic progenitor cells, 76% (+/-19.54 SD) compared to 64.19% (+/-29.77 SD). PrepaCyte-CB was also 12-fold more efficient in removing red blood cells and haemoglobin (P < 0.001) than Hetastarch. CONCLUSIONS These results show that PrepaCyte-CB offers superior separation of UCB when compared to Hetastarch.
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Affiliation(s)
- C Basford
- Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, UK
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Culture of embryonic-like stem cells from human umbilical cord blood and onward differentiation to neural cells in vitro. Nat Protoc 2008; 3:1046-55. [PMID: 18536651 DOI: 10.1038/nprot.2008.69] [Citation(s) in RCA: 63] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
This 3-week protocol produces embryonic-like stem cells from human umbilical cord blood (CBEs) for neural differentiation using a three-step system (cell isolation/expansion/differentiation). The CBE isolation produces a highly purified fraction (CD45-, CD33-, CD7-, CD235a-) of small pluripotent stem cells (2-3 microm in diameter) coexpressing embryonic stem cell markers including Oct4 and Sox2. Initial CBE expansion is performed in high density (5-10 millions per ml) in the presence of extracellular matrix proteins and epidermal growth factor. Subsequent neural differentiation of CBEs requires sequential introduction of morphogenes, retinoic acid, brain-derived neurotrophic factor and cyclic AMP. Described methods emphasize defined media and reagents at all stages of the experiment comparable to protocols described for culturing human embryonic stem cells and cells from other somatic stem cell sources. Neural progenitor and cells generated from CBEs may be used for in vitro drug testing and cell-based assays and potentially for clinical transplantation.
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