人脐血造血干细胞体外扩增方法初探  

Investigation of the effectiveness of in vitro expansion of human umbilical cord hematopoietic stem cells

作　　者：曲鑫建[1] 李惠侠[2] 丰美福[3] 孙世铎[2] 

机构地区：[1]右江民族医学院基础学院,广西百色533000 [2]西北农林科技大学动物科技学院,陕西杨凌712100 [3]中国科学院动物研究所生物膜与膜生物国家重点工程实验室,北京100080

出　　处：《基础医学与临床》2007年第9期1048-1052,共5页Basic and Clinical Medicine

基　　金：中国科学院基金(KSCX2-SW-322)

摘　　要：目的建立人造血干细胞(HSC)体外大量扩增的最佳培养模型。方法采用模拟骨髓微环境的成骨细胞共培养法、条件培养基及模拟微重力效应三维培养等方法进行人脐带血来源HSC体外扩增。结果在用含15%的条件培养基与成骨细胞共培养条件下,HSC的扩增倍数最高,第7天和第14天扩增倍数分别为6.46和14.82倍。在此条件下进行集落形成实验对扩增后造血干细胞多能性的功能检测发现,粒细胞-巨噬细胞集落形成单位(CFU-GM)及红系集落形成单位(BFU-E)数目也最多。结论以成骨细胞作为饲养层细胞,添加15%的条件培养基不仅能使HSC在体外大量增殖,并且具有维持HSC自我更新和多向分化潜能。Objective To investigate the effectiveness of in vitro expansion of human umbilical cord hematopoietic stem cells （HSCs） and to optimize the culture conditions. Methods HSCs were cocultured with osteoblasts in the three-direction （3D） culture system or condition medium （CM） to study the expansion effect on HSCs. Results In the conditions of cocultured with osteoblasts and added with 15% CM produced an expansion of the HSCs at a higher level, the HSCs were evaluated that expansion 6. 46-fold after 7^th days incubation while 14. 82-fold after 14^th days incubation. At the same conditions, the number of colony forming units of granulocyte and macrophage （CFU-GM） and erthroid burst-forming unit （CFU-E） were also significantly higher than that of other groups. Conclusion HSCs were cocultured with osteoblast in 15% CM medium not only achieves admirable results for in vitro expansion HSCs but also sustains the self-renew capacity.

关 键 词：造血干细胞 体外扩增 条件培养基 三维培养 

分 类 号：Q813.1[生物学—生物工程]