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CD34免疫亲合柱在脐血造血干 祖细胞分离与扩增中的应用 |
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王郡甫 田志刚 侯桂华 张建华 张彩 孙
摘 要 目的：观察CD34免疫亲合柱对脐血造血干/祖细胞分离纯化的 效果及纯化后CD34+细胞的增殖分化特性。方法：采用CD34免疫亲合柱分离脐血单个核细胞(MNC)中的CD34+细胞 ，流式细胞技术(FACS)进行细胞表面标志测定。将分离前后的细胞加入造血生长因子进行液 态扩增培养和多向祖细胞集落(CFU-GEMM)培养。结果：经CD34免疫亲合柱分离后脐血中CD34+细胞为49.62%±17.69 % ，明显高于脐血MNC(1.17%±0.68%)，细胞回收率为54.38%±11.91%。分离后CD34+细 胞 和脐血MNC经造血生长因子刺激培养20 d分别扩增561.00倍和44.44倍。培养至12 d时，免 疫亲合柱分离组CD34+细胞阳性率为53.38%，对照组为7.91%。分离组CFU-GEMM产率明 显高于对照组(P＜0.001)。结论：CD34免疫亲合柱应用于脐血造血干/祖细胞的分离可充分富集CD3 4＋细胞，且分离后的CD34＋细胞具有明显的增殖效应和CFU-GEMM形成能力。
关键词 CD34＋细胞 免疫亲合柱 扩增
中国图书分类号 R392-33
The application of CD34-immunoaffinity column in the i s olation and expansion of hematopoietic stem and progenitor cells of cord blood
WANG Jun-Fu,TIAN Zhi-Gang,HOU Gui-Hua et al.
(Shandong Academ y of Medical Sciences,Jinan 250062)
Abstract Objective:To observe the effect of CD34 immunoaf finity column on the isolation of hematopoietic stem and progenitor cells of cor d blood and the proliferatory and expansion characteristics of CD34＋ cells in vitro.Methods:CD34＋cells were isolated from cord blood using CD34 immunoaffinity column.Cell surface antigens were analysed by FACS.Th e separated and un-separated cells were cultured with human hematopoietic growt h factors(HGFS)in liquid culture system and CFU-GEMM culture system.Results:CD34＋ cells were enriched with a purity of 49.6 2%±17.69% and a recovery of 54.38%±11.91% using the immunoaffinity column.The sep arated CD34＋cells and cord blood MNC expanded to 561.00 folds and 44.44 folds respectively after cultured with HGFS for 20 days.The percentages of CD34＋ c ells in separated group and control were 53.38% and 7.91% respectively after cu ltured for 12 days.The number of CFU-GEMM in separated group was significantly higher than that in control(P＜0.001). Conclusion:Using CD34 immunoaffinity column the hematopoie tic stem and progenitor cells can be enriched efficiently and the separated CD3[1] [2] [3] 下一页
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