摘要
目的建立一株能在体外长期培养并具有多向分化潜能的人骨髓间充质干细胞(HBMSC).方法利用差异贴壁法从人肋骨中分离纯化HBMSC,在体外通过地塞米松、胰岛素诱导分化为脂肪细胞,用VitC、β-磷酸甘油、地塞米松诱导下可分化成骨细胞,裸鼠体内分化为软骨样细胞及成骨样组织,采用流式细胞仪检测细胞表面CD31、CD34、CD45、CD9、CD90、CD105和CD106.以2.5×107细胞接种于裸鼠皮下进行致瘤实验,用DY-NALREU-SSO反向杂交法检测HLA.结果细胞已连续培养超过30代仍能稳定生长,细胞表面特性为CD105、CD106阳性,而CD31、CD34、CD45、CD9、CD90为阴性.在体外能诱导成脂肪细胞、成骨细胞.无致瘤性,在裸鼠体内能形成软骨细胞及骨样组织.结论培养了一株能在体外长期培养及具有多向分化潜能的人骨髓间充质干细胞.
Purpose: To establish a kind of cell strain named human bone marrow mesenchymal stem cells HBMSC which are defined as multi-potential cells and can be cultured in vitro for long term. Methods: Using differential time adherent methods to isolate and purify the HBMSC from human rib. Fat cells was induced in medium supplemented with dexamethason and insulin. Osteogenic lineage was induced in medium supplemented with VitC, β-glycerophophate and dexamethason. The progeny of HBMSC gave rise to cartilage and skeletal tissues in nudemice model. The expression of cell surface markers were assessed using following monoclonal antibodies: CD31, CD34, CD45, CD9, CD90, CD105, CD106 by FACS. Nude mice were administrated 2.5 x 107 cell subcutaneously to verify the carcinogenicity. DYNAL RFU-SSO reversal hybridization were used to detect HLA. Results: The cell were cultured continuously over 30th passages and growed stablely. Phenotypic analysis indicated the strong expression of CD105, CD106 did no expression of CD34, CD45, CD9, CD90. HBMSC could be induced to fat cells, osteogenic cells in vitro. It could give rise to cartilage, skeletal tissues in nude mice nodel and without carcinogenicity. Conclusions: We established a kind of cell strain named human bone marrow mesenchymal stem cells in vitro which can be cultured for long term with multi-potent.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
2003年第3期232-234,F003,共4页
Fudan University Journal of Medical Sciences
基金
上海市卫生局科技发展基金(01433)
