脐带血CD34＋细胞体外扩增优化条件的研究

Study on optimized conditions for cord bloodCD34＋ ceils amplification in vitro

目的分析比较不同细胞饲养层、不同刺激因子、脐血CD34＋细胞的纯度和含量等因素对脐血CD34＋细胞在体外扩增的影响，以筛选其体外扩增的优化方案。方法采用正交实验，通过MTT法、细胞计数法及流式细胞分析技术测定脐血CD34＋细胞的增生情况；并采用以上方法及半固体培养体系对该细胞优化条件下的增生情况及集落形成能力进行检测。结果优化条件下脐血CD34＋细胞增生10倍，明显高于对照组的2．8倍（P〈0．01），集落形成（CFU—C36．67±6．11）也较对照组强（CFU—C16．33±1．53）（P〈0．01）。结论该培养系统能促进脐血CD34＋细胞体外扩增并能较好保持脐血CD34＋细胞的干细胞性质。

Objective To set up the optimized conditions,the amplification of cord blood CD34＋ cells in vitro were analyzed by comparing the conditions such as different feeder-layers, stimulating-factors or purity/contents of those cells. Methods The cord blood CD34＋ ceils proliferation was analyzed by the methods of MTr, cell counting, and flow cytometer. The amplification and clone-forming ability of cord blood CD34＋ cells were detected under optimized condition. Results The growth rates of cord blood CD34＋ cell under optimized conditions（10 times） were significantly higher than that of the control（2.8 times） （P 〈0.01）, and the cloneforming ability of cord blood CD34＋, cells under optimized conditions（CFU-C 36.67±6.11） were also better than that of the control（CFU-C 16.33±1.53） （P 〈0.01）. Conclusion The cord blood CD34＋ cells proliferation can be promoted in the co-cultured system, and the character of the stem cells were kept well in that system.