不同方法分离的大鼠骨髓间充质干细胞增殖活性和成脂分化能力的比较  

Comparison of the proliferation ability and adipose potential of rat bone marrow mesenchymal stem cells isolated by different methods

作　　者：庄淑波[1] 刘毅[1] 陈克明[1] 肖斌[1] 

机构地区：[1]兰州军区兰州总医院烧伤整形外科,甘肃兰州730050

出　　处：《中国美容医学》2007年第1期21-24,共4页Chinese Journal of Aesthetic Medicine

基　　金：全军"十一五"医学科研面上项目(06MA079);甘肃省自然科学基金(3zs061-A25-099)

摘　　要：目的:通过比较密度梯度离心和贴壁筛选两种方法所获MSCs的增殖活性和成脂分化能力,探索MSCs体外分离和纯化的较好方法。方法:应用密度梯度离心和贴壁筛选两种方法分离纯化骨髓MSCs,并通过MTT法检测其增殖活性,用第三代MSCs进行成脂诱导,以观察两种方法所获MSCs的成脂分化能力。结果:密度梯度离心法所获MSCs纯度高,呈纺缍形或小三角形,增殖快,约7～10天融合;贴壁筛选法分离的原代细胞中红细胞。破骨细胞等杂质细胞较多,增殖速度相对较慢,且经数次传代仍有较多杂质细胞存在,经成脂诱导后,两种方法所获MSCs表现出相似的成脂能力,油红O染色细胞计数值与光密度值比较两组无显著差异(P值>0.05)。结论:密度梯度离心法是体外分离和纯化MSCs较好的方法,所获细胞增殖活性高,与贴壁筛选法所获MSCs具有相似的成脂分化能力。Objective To explore a ideal method of isolation and purification of bone marrow MSCs in vitro by comparison of proliferation ability and adipose differentiation potential of the cells obtained by two different methods. Methods Density gradient centrifugation and plastic adherence methods were used for isolation of bone marrow MSCs from adult SD rats and the proliferation ability of the cells was analyzed by MTT examination. In order to observe the adipose differentiation ability of MSCs abtaind by two methods cells of passes 3 were indulsed. Results High purity of MSCs were obtained by gradient centrifugation. And the cells proliferated fast and got confluence in 7 to 10 days. Other cells like red blood cells and osteoctast, cells could be seen in primary MSCs obtained by plastic adherent method. These MSCs proliferated slow and even after several passages red blood cells and other cells could also be observed.But similar adipose differentiation ability,of MSCs obtaind by two methods showed after they were induced.No significant differences were found from Oil red O coloration and optical density. Conclusion Gradient centrifugation might be a ideal method for isolation and purification of MSCs from bone marrow. The cells showed better proliferation ability than that of those isolated by plastic adherent methods,meanwhile, two cells have the similar adipose differentation potential.

关 键 词：骨髓间充质干细胞 细胞培养 成脂分化 组织工程 

分 类 号：Q2-06[生物学—细胞生物学]