M-Reboredo N, Díaz A, Castro A, Villaescusa R G
Department of Cryobiology, Galician Blood Transfusion Center, Santiago de Compostela, Spain.
Bone Marrow Transplant. 2000 Dec;26(12):1263-70. doi: 10.1038/sj.bmt.1702728.
Umbilical cord blood (UCB) transplantation is being used as an alternative source of hematopoietic stem cells for bone marrow reconstitution. Separation and processing of UCB samples in large numbers for storage in cord blood banks ideally needs to be partially automated. The aim of this study was to establish and standardize a method for unrelated cord blood banking as well as the biological characterization of the samples. Up to October 1999, a total of 938 UCB units (mean volume 84.6 +/- 23.6 ml, nucleated cell (NC) count 0.90 +/- 0.37 x 109, total CFU-GM 79 +/- 72 x 104, CD34+ cell count 2.46 +/- 2.72 x 106) had been collected. Twenty-three per cent of all UCB samples had a NC count below 0.4 x 109 and were discarded. The initial bacterial contamination rate was reduced to less than 5% as a result of extensive training in collection procedures. Using a modification of a triple bag system and adding a solution of 6% hydroxyethyl starch, the UCB was separated by two centrifugation steps into three components: buffy coat, red cell and plasma fractions. The overall recoveries for NC, CFU-GM and CD34+ cells were 87.4 +/- 8.5%, 88.8 +/- 6.6% and 90.3 +/- 12.4%, respectively, in a mean final volume of 27 +/- 4.2 ml.
脐带血（UCB）移植正被用作造血干细胞的替代来源，用于骨髓重建。为了在脐带血库中储存，大量UCB样本的分离和处理理想情况下需要部分自动化。本研究的目的是建立并标准化一种无关脐带血库的方法以及样本的生物学特性鉴定。截至1999年10月，共收集了938个UCB单位（平均体积84.6±23.6毫升，有核细胞（NC）计数0.90±0.37×10⁹，总集落形成单位-粒细胞巨噬细胞（CFU-GM）79±72×10⁴，CD34⁺细胞计数2.46±2.72×10⁶）。所有UCB样本中有23%的NC计数低于0.4×10⁹，因此被丢弃。由于在采集程序方面进行了广泛培训，初始细菌污染率降至5%以下。采用改良的三袋系统并添加6%羟乙基淀粉溶液，通过两步离心将UCB分离为三个组分： Buffy 层、红细胞和血浆部分。在平均最终体积为27±4.2毫升的情况下，NC、CFU-GM和CD34⁺细胞的总体回收率分别为87.4±8.5%、88.8±6.6%和90.3±12.4%。
