人脐带间质干细胞分离纯化及基本生物学特性研究

目的探讨人脐带间质干细胞（MSC）的分离、纯化、扩增方法,研究其基本生物学特性.方法无菌条件下收集剖宫产新生儿脐带,分别用组织块贴壁法和酶消化法获取脐带间质干细胞,进行原代培养.应用DMEM/F12培养液进行纯化和扩增培养.用流式细胞仪检测MSC的细胞表面标志.绘制细胞生长曲线并测定细胞周期.结果两种分离方法均可获得MSC,原代培养12～14 d后可达90%融合,细胞可传代20代以上.流式细胞仪检测结果显示,脐带MSC强表达CD13、CD29、CD44、CD105,弱表达CD106,不表达CD34、CD11a、CD14、CD31、CD45.结论脐带MSC在体外有较强的增殖能力,可作为组织工程的种子细胞.

BIOLOGICAL STUDY OF HUMAN UMBILICAL CORD DERIVED MESENCHYMAL STEM CELLS

Objective To explore the isolation,purification and expansion(method) of human umbilical cord derived mesenchymal stem cells(UCMSC).MethodsThe umbilical cords from cesarean newborns were collected in sterile condition.Me-(senchymal) stem cells were obtained by enzyme digest method and cultured in DMEM/F12 medium.Surface antigens of UCMSC were detected by FACS.The cell growth curve and cell cycle of UCMSC were analyzed.ResultsMSCs could be obtained by both of the two methods.After 12-14 days of primary culture,isolated MSCs reached 90% confluence and the cells could expand at least 20 passages.FACS analysis showed that UCMSCs were positive for CD13,CD29,CD44,and CD105 and negative for CD106,CD34,CD11a,CD14,CD31,and CD45.ConclusionUmbilical cord derived MSCs has strong proliferation capacity,which can be used as the seed cells for tissue engineering.