从脐带血中提取的CD34阳性干细胞在冷冻保存6个月后仍可存活

M. Muhibi, V. O. Mabayoje, J. Komolafe
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引用次数: 0

摘要

脐血可作为骨髓移植的替代来源,其用途正在发展成为一个治疗血液系统疾病、免疫缺陷和特定遗传疾病患者的新领域;包括血红蛋白病。目的是评估冷冻脐带血作为HSC来源的可行性,该来源可能适合移植。从30名同意的母亲的脐带上采集血样,并将其分装到5个装有甘油的冷冻瓶中,在-20°C下冷冻;同时通过使用CD34作为HSC的标记物的免疫表型在新鲜柠檬酸盐样品上进行定量测定。Partec Cyflow立方体6用于在用从Sysmex Partec获得的特异性荧光色素/抗体标记细胞后测量活细胞。连续5个月以一个月为间隔进行重复定量,并使用T独立检验分析产生的结果。连续6次计数的平均值±标准误差(SEM)分别为20798±2750、19849±2691、19223±2637、18363±2582、17052±2583和16184±2423。将冷冻保护的样品与基线进行比较时获得的p值分别为0.806、0.681、0.521、0.325和0.213;反映出随后的计数与基线计数无显著差异。因此,在资源匮乏的环境中,将干细胞储存在冷冻保护剂中并在-20°C下冷冻长达6个月是一种安全的选择,而不会显著降低生存能力。应该探索这种替代方案,并进行进一步的研究,有可能延长月数。关键词:CD34+细胞,免疫表型,干细胞,冷冻保存。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
CD34 positive stem cells recovered from cord blood remain viable after six months of cryoprotective storage process
Cord blood can be used as an alternative source for bone marrow transplantation and its use is developing into a new field of treatment for patients presenting with haematological disorders, immunological defects and specific genetic diseases; including haemoglobinopathies. The aim was to assess the viability of frozen cord blood as a source of HSC which may be suitable for transplantation. Blood specimens were obtained from umbilical cords of 30 consenting mothers and dispensed into 5 cryovials with glycerine for freezing at -20°C; while quantitative assay was carried out on a fresh citrated sample by immunophenotyping using CD34 as marker of HSC. Partec Cyflow cube 6 was used to measure viable cells after labelling the cells with specific fluorochrome/antibody obtained from Sysmex Partec. A repeat quantification was carried out at one month interval for 5 consecutive months and results generated were analysed using T- independent test. The mean ± standard error of mean (SEM) for the 6 consecutive counts were 20,798±2750, 19849±2691, 19223±2637, 18363±2582, 17052±2583 and 16184±2423. The p values obtained when the cryoprotected samples were compared to the baseline were 0.806, 0.681, 0.521, 0.325 and 0.213; reflecting that subsequent counts were insignificantly different from the baseline count. Thus, it is a safe alternative in resource-poor setting to store stem cells in a cryoprotective agent and freeze at -20°C for up to 6 months, without significant depreciation in viability. This alternative should be explored and further researches should be conducted with possibility of extending the number of months. Key words: CD34+ cells, immunophenotyping, stem cells, cryopreservation.
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