脐血造血细胞体外维持和扩增的实验研究  

The Experimental Study on ex vivo Maintenance Expansion of Hematopoietic Cells from Cord Blood

作　　者：王红卫[1] 缪竞诚[2] 杨吉成[2] 盛伟华[2] 李丽娥[2] 彭明发[2] 查纪坤 

机构地区：[1]复旦大学 [2]苏州医学院

出　　处：《复旦学报（自然科学版）》2000年第3期247-250,共4页Journal of Fudan University：Natural Science

基　　金：江苏省科委资助项目 (DS 980 37)

摘　　要：探讨胎儿、儿童、成人骨髓基质细胞对脐血造血细胞体外长期生存的维持能力 ,以及细胞因子组合在基质接触和无基质的培养体系中扩增脐血干、祖细胞的效应 .将连续二次Ficoll Hypaque分离的脐血单个核细胞(MNCs)接种至经辐照预处理的各种骨髓基质细胞上 ,持续培养 6周 ,定期检测祖细胞 (CFC)和长期培养起始细胞 (LTC IC) ;使用干细胞因子 (SCF)、白细胞介素 (IL) 3、IL 6组合 ,在基质接触和无基质体系中 ,扩增脐血干、祖细胞 .结果表明 :①成人骨髓基质细胞能更有效地维持脐血 2周、4周CFCs的数量 ,在 10 5初始接种MNCs中 ,产量分别为 6 4 5± 11 3和 43 6± 7 6 ;而胎儿骨髓基质细胞对脐血LTC ICs表现出更强的支持作用 ,其LTC ICs的产量为 18 5± 5 2 .②成人基质细胞接触的扩增体系 ,CFCs扩增最为显著 ,其CFCs的产量为扩增前的 8 4倍 ;胎儿基质接触的扩增体系能够更有效地扩增LTC ICs ,其产量为扩增前的 3 9倍 .基于上述结果 ,认为不同发育阶段的骨髓基质细胞对脐血CFCs和LTC ICs的生存 ,表现出不同的维持效应 ;基质接触的扩增体系更适合于脐血原始造血细胞的扩增 .The potential of fetal, child and adult bone marrow stromal cells to ex vivo maintain the long term survival of cord blood hematopoietic cells, as well as the expansion effect of the cytokine combination were investigated. Mononuclear cells (MNCs) from cord blood were seperated by the continiously double Ficoll Hypaque method, then cultured on the various preestablished irradiated stromal layers for six weeks, the number of progenitors(CFCs) and long term culture initiating cells(LTC ICs) were meassured at different time point. In addition, the combination of stem cell factor (SCF), interleukine 3(IL 3), IL 6 was used to expand stem cells and progenitors in stroma contact or stroma free system. As a results, it is found that, first, the adult bone marrow stromal cells were more efficient to maintain the 2 and 4 weeks CFCs from cord blood with the production 64.5±11 3 and 43.6±7.6 respectively, and fetal stromal cells were more efficient to support the survival of LTC ICs with the production 18.5±5.2. Secondly, the more significant expansion of CFCs was found in adult stroma contact expansion system with 8.4 fold increase as compared with initial value; the more remarkable increase of LTC ICs was obtained in the fetal stroma contact expansion system with 3.9 fold increase. In a word, the results indicate various stromal cells from different developmental stage displayed different effects on the maintenance of cord blood CFCs and LTC ICs, and the stroma contact expansion system may be more suitable for the amplification of the primitive hematopoietic cells in cord blood.

关 键 词：造血细胞 骨髓基质细胞 体外扩增 脐血 体外维持 

分 类 号：R331.2[医药卫生—人体生理学]