人骨髓间充质干细胞联合细胞因子的无血清培养体系体外扩增脐血干细胞研究  

Investigation of the effectiveness of ex vivo expansion of human cord blood stem cell

作　　者：费小明[1] 吴雨洁[1] 唐宇宏[1] 沈文怡[1] 陆化[1] 李建勇[1] 汪承亚[1] 

机构地区：[1]南京医科大学第一附属医院,210029

出　　处：《江苏医药》2004年第4期244-246,I002,共4页Jiangsu Medical Journal

基　　金：南京医科大学创新基金 (CX2 0 0 10 0 4)

摘　　要：目的　探讨以人骨髓间充质干细胞 (BM MSC)为基质的无血清培养方法 ,体外扩增脐血造血干细胞 (CBSC) ,研究BM MSC支持造血的功能和体外扩增时间的选择。方法　用含血小板生成素 (TPO)、干细胞因子 (SCF)、flt3/flk2配体 (FL)和粒细胞 集落刺激因子 (G CSF)的无血清培养液 ,以人BM MSC为基质 ,比较体外扩增脐血CD34+ 细胞 7d及 14d后总细胞 (TC)、CD34+ 细胞和集落形成单位 (CFU)数增加倍数。结果　在以人BM MSC为基质及TPO、FL、SCF和G CSF这四种细胞因子作用下 ,经体外扩增 7d后TC、CD34+ 细胞、CFU GM和CFU C数较起始分别增加了87、16、15和 2 6倍 ;14d后较起始分别增加了 4 2 7、38、12 5和 10 4倍 ,与 7d扩增倍数有显著性差异。结论　以人BM MSC为基质的无血清体外培养体系可以有效扩增CBSC ,有潜在的临床应用价值 ,扩增时间应以Objective To investigate the effectiveness of ex vivo expansion of human cord blood stem cell (CBSC) using serum-free system and co-culture with human bone marrow derived mesenchymal stem cell(BM-MSC),and also to optimize the culture duration.Methods CD34 + CBSC were cocultured with BM-MSC in the serum-free media containing thrombopoietin(TPO),flt3/flk2 ligand (FL),stem cell factor (SCF)and granulocyte-colony stimulating factor (G-CSF).The total cells (TC),CD34 + cells and colony forming units(CFU)of the expanded cells were evaluated on the 7th day or 14th day of culture.Results In the presence of both BM-MSC and four cytokines mentioned above,CBSC produced an expansion of 87-fold in TC,16-fold in CD34 +,15-fold in CFU-C and 26-fold in CFU-GM after 7-days' incubation,while CBSC produced an expansion of 427-fold in TC,38-fold in CD34 +,125-fold in CFU-C and 104-fold in CFU-GM after 14-days' incubation.There is significant difference between the two groups in TC,CD34 + and CFU expansion.Conclusion Our serum-free and co-culture system was shown to achieve admirable results for ex vivo expansion of CBSC,and may probably be employed for clinical application.Additionally,14 days' incubation is more efficient than the 7-day's culture in ex vivo expansion of CBSC.

关 键 词：骨髓间充质干细胞 细胞因子 无血清培养体系 体外扩增 脐血干细胞 造血干细胞移植 

分 类 号：Q813.11[生物学—生物工程] R457.7[医药卫生—治疗学]