Optimal processing of human umbilical cord blood for clinical banking.
Optimal processing of human umbilical cord blood for clinical banking.
复制标题
用于临床银行的人脐带血的优化处理。
作者：
P. Denning;C. Donaldson;A. Nicol;B. Bradley;J. Hows
人脐血(UCB)已成功地作为异基因造血干细胞的替代来源用于儿童移植。UCB的临床银行需要减少体积和耗尽红细胞以实现经济高效的存储。比较了Ficoll法、Percoll法、甲基纤维素法、明胶法、淀粉法和红细胞裂解法处理UCB的效果。由于单个脐带血标本在集落形成细胞和CD34+细胞含量方面差异很大，每个脐带血标本(n=26)均采用三种或三种以上技术处理，同时采用Ficoll作为“标准”方法。明胶对CFCs(92%)和CD34+细胞(86%)的回收率一直很高。明胶处理的脐血白细胞中CD34+占0.10-2.50%，批内变异为2.1%。结合个别实验数据，CD34+与氯氟化碳含量的相关性良好(r=0.77)。就CD34+和氟氯化碳回收率而言，Lysis排名第二，但由于涉及的数量很大，因此不太实用。Ficoll和Percoll位居第三，但它们的成本更高，技术也更复杂。淀粉的沉积被证明是缓慢的，而甲基纤维素处理失去了超过60%的氟氯化碳和CD34+细胞。经过明胶处理后，我们计算出70毫升脐带血的平均+/-SD为9+/-2×10(8)有核细胞，32+/-18×10(5)CD34+细胞，20+/-12×10(5)CFC95%以上的红细胞耗尽率。最近发表的计算机研究表明，在成人移植受者中，可能只需要2×10(5)CD34+细胞就可以持续植入异基因骨髓。我们得出结论，平均70毫升的脐带血捐献含有足够的骨髓再生细胞供成年受者使用。
Human umbilical cord blood (UCB) has been successfully used as an alternative source of allogeneic hematopoietic stem cells for pediatric transplantation. Clinical banking of UCB requires volume reduction and red cell depletion for cost-effective storage. We have compared processing UCB by Ficoll, Percoll, methylcellulose, gelatin, starch, and red cell lysis. As individual UCB collections vary widely in colony forming cell (CFC) and CD34+ cell content, each UCB (n = 26) was processed by three or more techniques in parallel with Ficoll as the "standard" method. Gelatin gave a consistently high recovery of CFC (92%) and CD34+ cells (86%). Between 0.10-2.50% of the leukocytes in gelatin-treated UCB were CD34+ with an intra-assay variation of 2.1%. Combining data from individual experiments, the correlation between CD34+ and CFC content was excellent (r = 0.77). Lysis rated second in terms of CD34+ and CFC recoveries but is not as practical because of the large volumes involved. Ficoll and Percoll came third but are more expensive and more involved techniques. Starch sedimentation proved to be slow, while methylcellulose processing lost over 60% of CFC and CD34+ cells. After gelatin processing, we calculated 70-mL donations of UCB would contain a mean +/- SD of 9 +/- 2 x 10(8) nucleated cells, 32 +/- 18 x 10(5) CD34+ cells, and 20 +/- 12 x 10(5) CFC with greater than 95% red cell depletion. Recent published computer studies suggest that as few as 2 x 10(5) CD34+ cells may be needed for sustained engraftment of allogeneic marrow in adult transplant recipients. We conclude that average 70-mL UCB donations contain sufficient marrow repopulating cells for adult recipients.