小鼠骨髓间充质干细胞(BMSCs)的体外分离扩增及鉴定  

The Isolation and Identification of Mouse Bone Marrow Mesenchymal Stem Cells(BMSCs) in Vitro

作　　者：石云[1] 张晓英[2] 

机构地区：[1]长治医学院附属和济医院儿科,046011 [2]北京军区总医院附属八一儿童医院

出　　处：《长治医学院学报》2012年第3期164-166,170,共4页Journal of Changzhi Medical College

摘　　要：目的:探讨一种简单易行的的小鼠骨髓间充质干细胞(bone marrow mesenchymal stemcells,BMSCs)体外分离培养方法。方法:通过全骨髓贴壁分离法体外分离、扩增小鼠BMSCs。观察原代及传代小鼠BMSCs形态变化,对第3代小鼠BMSCs表面抗原CD34、CD45、CD105和CD106进行流式细胞仪检测。结果:小鼠BMSCs的原代接种培养24h后可见大量悬浮细胞,少许贴壁细胞,呈小圆形;72h后贴壁细胞逐渐增多,培养至第7天细胞伸展成长梭形,呈集落生长。BMSCs在培养的2～6d细胞增殖较慢,7～10d细胞增殖较快。传代后的小鼠BMSCs 24h基本全部贴壁,呈均匀性梭行细胞样生长。流式细胞仪检测结果显示:CD105、CD106表达阳性,表达率分别为92.2%、90.4%;CD34、CD45表达阴性,表达率分别为8.8%、13.1%。结论:采用全骨髓贴壁分离法操作简便,是培养小鼠BMSCs的理想方案。流式细胞术可以鉴定体外分离培养的小鼠BMSCs。Objective：Establish the method of isolation and identification of the mouse bone marrow mesenchymal stem cells （BMSCs） in vitro. Methods：Cultured mouse BMSCs with the simple and available method of adherent cells separation, Microscope were performed to observe the changes of cell morphology during this procedure. Cells were analyzed with a FACS can for CD34, CD45, CD105 and CD106 expression. Results： After several discarding supernatant （24 -72 h）, the percent of adhered ceils grew more,and there were colony formation were seen. the cell appeared long spindle. At the first 2-6 days, the cells growed very slow, however, cells growed faster in 7-10 days, which were ＂whirlpool-like＂. BMSCs extracted from the BM formed a homogeneous population of cells by the second passage. The analysis of cell surface phenotype indicated that the BMSC population was positive for CD105, CD106, whereas negative for CD34 and CD45, with more then 90% uniformity. Conclusion=The method was very simple,convenient and useful, by which mouse BMSCs were successfully isolated and identified.

关 键 词：骨髓 间充质干细胞 小鼠 流式细胞术 全骨髓贴壁分离法 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]