人脐带血间充质干细胞体外分离培养及生物学特性研究  

Study on Isolation and Culture of Mesenchymal Stem Cells from Human Umbilical Cord Blood in Vitro and Their Biological Characteristics

作　　者：程文广[1] 罗高兴[1] 黄正根[1] 贺伟峰[1] 袁顺宗[1] 陈希炜[1] 吴军[1] 

机构地区：[1]第三军医大学西南医院烧伤研究所重庆市疾病蛋白质组学重点实验室,重庆400038

出　　处：《现代生物医学进展》2008年第3期424-426,共3页Progress in Modern Biomedicine

基　　金：军队"十一五国际合作项目(06H031);军队十一五专题(06-Z-054);重庆市自然基金(2005BB5044);校院基金(SWH2005A010)

摘　　要：目的:探讨并建立稳定可靠的人脐带血间充质干细胞(human umbilical cord blood mesenchymal stem cells,hUCBMSCs)体外分离和培养方法。方法:采用密度梯度离心法分离脐带血单个核细胞,利用MSC易黏附塑料贴壁生长的特性,分离MSC并进行体外扩增培养,观测其形态学特征,绘制不同代数hUCBMSc生长曲线,流式细胞技术检测细胞表面标志物CD29、CD34、CD105、HLA-DR等表达情况。结果:本法可成功、可靠地在体外自hUCB中分离培养出粘附细胞,该细胞在体外培养呈梭形或成纤维样,其指数生长倍增时间约为36h,其细胞表面表达β1整联蛋白CD29、CD105(endoglin),不表达造血细胞标志物CD34以及HLA-DR。结论:人脐带血中存在间充质干细胞,采用优化的实验方法可以提高培养成功率,稳定获得MSC并体外扩增培养。人脐带血可以作为获得间充质干细胞稳定充足的来源。Objective： To establish an Optimized method to isolate, culture and expend mesenchymal stem cells （MSCs） from human umbilical cord blood （hUCB） in vitro. Methods： The mononuclear cell was isolated by density gradient centrifugation with lymphocyte separation medium from hUCB collected from full term deliveries scheduled for cesarean section. The MSCs were purified by adhesion with acidized D-MEM/F-12 medium. After that, the growth curves of MSCs in different passages were drawn by cell counting, and the expressions of CD29, CD105, CD34, HLA-DR in MSCs was detected by flow cytometry. Results： The adhesion cells could be obtained successfully and reliably from hUCB. The cells displayed a fibroblast-like morphology, and several MSCs-related antigens such as CD29 and CD105 were expressed, while neither CD34 nor HLA-DR could be found on the cells by FACS. Conclusion： MSCs could be obtained from hUCB with the method mentioned in this article and hUCB could be an alternative source of MSCs for experimental and clinical applications.

关 键 词：人脐带血 间充质干细胞 分离培养 生物学特性 

分 类 号：Q813[生物学—生物工程]