不同细胞因子组合对脐血造血干细胞体外扩增效果分析  

Effect of different human recombinant cytokine combinations on the expansion of cord blood haemopoietic stem cells

作　　者：孟庆宝 魏亚明[2] 张印则[3] 

机构地区：[1]深圳市人民医院,广东深圳518020 [2]广州医学院附属广州市第一人民医院 [3]深圳市输血医学研究所

出　　处：《中国输血杂志》2009年第6期443-446,共4页Chinese Journal of Blood Transfusion

基　　金：深圳市科技局资助项目;项目编号:No200702107

摘　　要：目的观察不同细胞因子组合对脐血干细胞体外扩增效果的影响。方法用含15%胎牛血清的改良细胞培养液,按加入的不同细胞因子组合分成7个实验组(A—G)进行细胞培养。分别于培养0d、3d、7d、14d取培养液,台盼蓝染色计数单个核细胞的总数、流式细胞仪分析CD34+细胞含量。结果与对照组相比,含细胞因子的7个实验组单个核细胞的数量均显著增加,除F组在培养d14时扩增达到峰值(15.60±10.90)×109/L外,其它各实验组在扩增d7时,均达到了扩增峰值并以E组为最高(17.11±6.12)×109/L,培养延续到d14时,各组均有不同程度的下降。所有实验组CD34+细胞的百分比也显著提高,A、D、E、F、G组均在d7达到了峰值,B、C组则在第14d达到峰值。经计算,扩增7d时CD34+细胞单位体积含量范围在(34.7±10.4)×107/L(A组)—(168.6±43.5)×107/L(这种表述似乎不太合适),对应的扩增达10—50倍。结论不同细胞因子组合可以提高脐血单个核细胞和CD34+细胞的扩增效率,SCF+IL-3,-6,-2,-4的组合在d7时达到了最佳的扩增。Objective To study the expansion efficiency of cord blood haemopoietic stem cells when cultured with different haman recombinant cytokine combinations. Methods Mononuclear cells separated from cord blood were cultured in Iscove Modified Dulbecco＇s medium（IMDM）. Seven experimental groups （A-G） were set up according to the addition of different cytokine combinations. Mononuclear cells were counted by trypan blue exclusion and the CD34^＋ cells were sorted by flow cytometry on the 3^rd, 7^th, and 14^th culture day. Results Compared with the control group, the amount of monunuclear cells in each experimental group increased significantly. Except group F, which reached the peak expansion of （15.60 ±10.90）×10^9/L, on the 14^th culture day, all other six groups showed peak expansion on the 7th day, among which group E was the highest [（17. 11±6. 12） ×10^9/L]. The percentage of CD34^＋ cells in group A, D, E, F, and G peaked on the 7^th culture day, and the peak expansion in group B and C was observed on the 14^th culture day. The number of CD34^＋ cells in unit volume among the seven groups ranged from （34. 7 ± 10. 4） × 10^7/L （group A） to （ 168.6 ± 43.5） ×10^7/L （group E）, with 10-50 folds of expansion. Conclusion All the experimental groups with different human recombinant cytokine combinations could improve the expansion efficiency of cord blood mononuclear cells and CD34^＋ cells. The group with the combination of SCF ＋ IL-3,-6,-2,-4 achieved the peak expansion on the 7^th culture day.

关 键 词：脐血 造血干细胞 单个核细胞 CD34^+细胞 扩增 

分 类 号：R331.1[医药卫生—人体生理学]