Donaldson C, Denning-Kendall P, Bradley B, Hows J
University of Bristol, Division of Transplantation Sciences, Southmead Hospital, Bristol, UK.
Bone Marrow Transplant. 2001 Feb;27(4):365-71. doi: 10.1038/sj.bmt.1702810.
Expansion of haemopoietic stem cells is proposed to combat graft failure in adult recipients following cord blood (CB) transplantation. Cultures are traditionally performed in medium containing FCS, but to transfer expansion to the clinic, 'good manufacturing practice' (GMP) standards are required. This study evaluated expansion cultures in culture bags and serum-free (SF) conditions, to comply with GMP, by analysing sub-populations of CD34(+) cells, colony-forming cells (CFC) and long-term culture initiating cells (LTC-IC). CD34(+)cell analysis has previously been used to measure clonogenic capacity and the CD34(+)CD38(neg) surface phenotype to measure primitive cell numbers. In this study, comparison of expansion in serum-replete medium with that in SF conditions demonstrated a lack of expression of CD38 on CD34(+) cells in the absence of serum. These findings must be considered in clinical studies using in vitro expansion in SF conditions, and the CD34(+)CD38(neg) phenotype should not be used to confirm maintenance, or expansion, of primitive progenitor cells.
有人提出，造血干细胞的扩增可用于对抗成人脐带血（CB）移植受者的移植物失败。传统上，培养是在含有胎牛血清（FCS）的培养基中进行的，但为了将扩增技术应用于临床，需要符合“良好生产规范”（GMP）标准。本研究通过分析CD34(+)细胞亚群、集落形成细胞（CFC）和长期培养起始细胞（LTC-IC），评估了在符合GMP的培养袋和无血清（SF）条件下的扩增培养。CD34(+)细胞分析以前曾用于测量克隆形成能力，而CD34(+)CD38(neg)表面表型用于测量原始细胞数量。在本研究中，将富血清培养基中的扩增与无血清条件下的扩增进行比较，结果表明在无血清的情况下，CD34(+)细胞上缺乏CD38的表达。在使用无血清条件下的体外扩增进行的临床研究中，必须考虑这些发现，并且CD34(+)CD38(neg)表型不应被用于确认原始祖细胞的维持或扩增。
