脐血造血干／祖细胞的密度分离实验研究

目的：为建立脐血造血细胞库，减少脐血储存空间，探讨脐血造血干／祖细胞的密度分离效果。方法：选用五种不同比密的Ficoll分离液（1.084、1.077、1.072、1.064、1.054g/L）分离脐血细胞，采用流式细胞技术（FCM）和全自动血细胞分析仪分析了41例脐血造血干／祖细胞和有核细胞（NC）的回收率并作了对比研究。结果：1.064g/L Ficoll分离脐血有核细胞回收率3.6％，淋巴细胞去除率为86.6％；所分离的单个核细胞(MNC）中CD34^ 细胞平均纯度为10.3％，最高可高达46.6％；所分离的细胞体外易形成CFU－GM及BFU－E。结论：不同比密分离脐血造血干／祖细胞具有不同影响。其中1.064g/L Ficoll分离脐血造血干／祖细胞是一种方便、实用、有效的方法。所分离的细胞体外有较强的增殖能力。

Experimental studies on density separation of hematopietic stem/progenitor cells from cord blood

Objective To investigate the effect of density separation of hematopoietic stem/progenitor cells(HSCs) for cord blob banking with storage volume reduction. Methods The separation efficiency of HSCs from cord blood were evaluated using five different densities Ficolls (1. 084, 1 .077,1. 072, 1 .064, 1. 054 g/L). The recovery of nucleated cells(NCs) and HSCs from 41 cord bloods were analyzed by flow Cytometry and blood autoanalyser. Results The recovery of NCs in 1 .064 g/L Ficoll spearated samples was 3. 6% with 86.6% lymphocyte depletion,The purity of CD34 +,the highest one reached 46.6%,was 10.3% on average in mononu clear cells(MNCs),The 1 .064 g/L Ficoll separated cells was easy to form colonies of CFU-GM and BFU-E in vitro. Conclusion The separation of hematopoietic stem/progenitor cells from cord blood was affected by different density Ficoll. The 1 .064 g/L Ficoll separation method was a convennient, practical, efficient method. The separated cells contained a higher percentage of HSCs in MNCs, which was demonstrated to have positive proliferation activity in vitro.