人脐血间充质干细胞的改良分离培养  

Improved method for isolation and culture of mesenchymal stem cells from human umbilical cord blood

作　　者：何晓婷[1] 李怀芳[1] 王建军[1] 孙向华[2] 

机构地区：[1]同济大学附属同济医院妇产科,上海200065 [2]同济大学附属同济医院中心实验室,上海200065

出　　处：《同济大学学报（医学版）》2012年第2期33-37,共5页Journal of Tongji University(Medical Science)

基　　金：上海市科学技术委员会科研计划项目(09411963700)

摘　　要：目的探讨人间脐血来源的充质干细胞在体外分离培养与扩增的方法。方法用重力离心-密度梯度离心法结合贴壁筛选法分离细胞,用含15%FBS的DMEM低糖培养基培养并传代,流式细胞仪检测P3代间充质干细胞的表面标志。结果脐血来源的单个核细胞24 h即开始贴壁,主要表现为破骨样细胞和间充质样细胞。经自然纯化法传代后,可得到形态单一的长梭形间充质干细胞。流式细胞仪检测结果显示该细胞表达CD44。结论脐血来源的MSCs可以在体外分离培养和扩增,细胞表面标记与骨髓来源间充质干细胞一致。Objective To improve the method for isolation and culture of human umbilical cord blood-derived mesenchymal stem cells（hUCB-MSCs） in vitro.Methods The cord blood mononuclear cells were isolated by density gradient centrifugation and directly adherence growth method.After cultured with DMEM containing low glucose and 15% FBS medium for 3 passages,surface antigen expression of MSCs was detected by flow cytometry.Results The cord blood mononuclear cells began to adherent to flasks in 24h,mainly displayed osteoblast-like cells and mesenchymal-like cells.By natural purification for 3 passages,long spindle shaped-MSCs were harvested.Test results from flow cytometry showed that the cells expressed CD44 strongly.Conclusion With the improved method hUCB-MSCs can be isolated,cultured and expanded in vitro.The harvested hUCB-MSCs express the identical surface antigen as human bone marrow-derived mesenchymal stem cells（BM-MSCs）.

关 键 词：人脐血 间充质干细胞 分离 培养 

分 类 号：R318[医药卫生—生物医学工程]