建立ICR小鼠骨髓间充质干细胞的体外分离培养及鉴定方法  

Establishment of the protocol for isolation,cultivation and identification of ICR mice bone marrow-derived mesenchmal stem cells in vitro

作　　者：杨玉彦[1,2] 王录美 孙铭学[1] 裴志鹏 冯雍炜 赵杰[1] 肖凯[1] 

机构地区：[1]第二军医大学热带医学与公共卫生学系防化医学教研室,上海200433 [2]第二军医大学干细胞研究中心,上海200433 [3]第二军医大学长海医院临床实验中心,上海200433

出　　处：《药学服务与研究》2016年第2期90-93,共4页Pharmaceutical Care and Research

基　　金：国家科学技术重大专项子课题(2013ZX09J13103-02C)

摘　　要：目的:建立一种简单的高效分离、培养、纯化和鉴定ICR小鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)的方法,初步研究其相关生物学特性。方法:全骨髓贴壁法分离培养BMSCs,用倒置显微镜观察细胞的形态和生长情况;用流式细胞仪鉴定细胞表面标记物;诱导细胞定向分化为成脂细胞、成骨细胞,以检测细胞多向诱导分化能力。结果:利用该方法纯化的BMSCs形态均一,多为梭形。第3代BMSCs进行流式检测CD29、CD44均呈阳性表达,表达率分别为97%、90%;CD34、CD45呈阴性表达;且能够成功诱导分化成成骨细胞、成脂细胞。结论:利用全骨髓贴壁法可以高效分离、纯化ICR小鼠的BMSCs,生物学特性稳定。Objective：To establish a simple and efficient method for isolation,cultivation,purification and identification of ICR mice bone marrow-derived mesenchymal stem cells（BMSCs）,and to make a preliminary study of its biological characteristics.Methods：BMSCs were isolated and cultured by whole bone marrow adherence method.The morphology and growth of cells were observed by invert-microscopy,and the membrane antigens of BMSCs were detected by flow cytometry.The proliferation and induction differentiation of BMSCs were identified by whether they had potencies of differentiating into adipogenic cells and osteoblasts.Results：The BMSCs purified by this protocol were homogeneously in the shape of fusiform.The third generation of BMSCs tested were positive for CD29 and CD44,with the expression rates of 97% and 90%,respectively.However,they were negative for CD34 or CD45.They could differentiate into adipogenic cells and osteoblasts.Conclusion：The whole bone marrow adherent culture method could effectively isolate,cultivate and identify BMSCs in mice with stable biologic features.

关 键 词：骨髓间充质干细胞(BMSCs) ICR小鼠 全骨髓贴壁法 

分 类 号：R965.2[医药卫生—药理学]