人脐带血间充质干细胞的体外培养及影响因素

目的:建立人脐带血的间充质干细胞(MSCs)体外培养、扩增的方法。方法:无菌条件下取正常足月的脐带血,分离单个核细胞,以含5%胎牛血清的低糖DMEM作为培养基进行培养和纯化获得贴壁细胞,测定MSCs的生长曲线,用流式细胞技术分析细胞的表面抗原。结果:脐血中的单个核细胞在适当的条件下分离、培养,获得的MSCs均一稳定地表达相关的抗原标记CD29、CD44、CD105,但不表达CD34、CD45、CD106和HLA-DR。结论:脐带血中含有的MSCs可在体外培养、扩增,能够作为间充质干细胞的一种有效来源。

In Vitro Cultured and Influencing Factor of Mesenchymal Stem Cells Derived from Human Umbilical Blood

Objective:To establish the method of in vitro culture and expansion of human umbilical blood-derived mesenchymal stem cells(MSCs) and to investigate their biological characteristics.Methods:HUCB were collected from full term deliveries csheduled for cesarean section,the cord bolld mononuclear cell were isolated by lymphocyte separation medium, purified and culture with DMEM-LG medium to produce adherent layer, the growth curves of MSCs were drawn, the surface antigenic features were analyzed by flow cytometry technique.Results:The HUCB-derived mononuclear cells, when seed in suitable medium, gave rise to adherent cells with the MSCs displayed a fibroblast-like morphology and expressed several MSCs-related antigens CD29,CD44,CD105,but did not express antigens CD34,CD45,CD106,HLA-DR,which was identical to human bone marrow-derived MSCs.Conclusions:MSCs in HUCB can be cultured and expanded in vitro,and could be regarded as an alternative source of MSCs for experimental and clinical applications.