大鼠骨髓间充质干细胞的培养、扩增与鉴定  

The Culture, Proliferation and Identification of Rat Bone Marrow-Derived Mesenchymal Stem Cells

作　　者：冯斌[1] 杨庭树[1] 梁丽玲[1] 

机构地区：[1]解放军总医院心内科,北京100853

出　　处：《天津医药》2009年第5期400-402,I0005,共4页Tianjin Medical Journal

基　　金：全军"十五"重点课题资助项目(项目编号:01Z036)

摘　　要：目的:建立大鼠骨髓间充质干细胞(MSCs)分离及培养的方法,探讨体外培养MSCs的生物学特性。方法:采用密度梯度离心法和贴壁筛选法纯化MSCs,镜下连续观察细胞的形态变化。流式细胞仪鉴定其表面抗原CD29、CD34、CD44和CD45的表达情况。结果:原代MSCs呈集落状生长,细胞呈梭形、纺锤形,呈放射状排列。传代后大都均匀分布生长,细胞生长迅速,倍增时间约45.5h。流式细胞术鉴定表明,培养的第3代和第5代MSCsCD44、CD29阳性,CD45和CD34阴性。结论:采用密度梯度离心法结合贴壁筛选法可成功分离和培养大鼠的骨髓MSCs,MSCs可以作为组织工程中种子细胞的来源。Objective： To establish a method of isolating and cultivating rat bone marrow mesenchymal stem cells （MSCs） in vitro and the biological characteristics thereof. Methods： Rat MSCs were isolated by density gradient centrifugation and cultured with L-DMEM. The morphology of MSCs was observed in primary and passage cuhure under inverted microscope. The cell surface antigens of CD29, CD34, CD44 and CD29 were detected through flow cytometer. Results： Cultured primary MSCs were spindle-shaped and had a typical fibroblast-like morphology, proliferated in colonies. After a series of passages, the attached cells became morphological homogeneous and proliferated quickly. During the log phase of growth, cells proliferated with a population doubling time of 45.5h. The surface marker identification showed that CD44 and CD29 were positive, but CD34 and CD45 were negative. Conclusion： The bone marrow MSCs can be successfully isolated, cultured and purified from rat bone marrow through density gradient centrifugation, adherent property and subculture. The MSCs can be used as a potential cell source for tissue engineering.

关 键 词：骨髓 间质干细胞 细胞培养技术 细胞增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R329-33[医药卫生—基础医学]