应用全骨髓贴壁法获取高纯度大鼠骨髓间充质干细胞的实验研究  

Experimental study on obtaining high purity of rat mesenchymal stem cells by whole bone marrow adherent culture method in vitro

作　　者：卢宁[1] 赵龙凤[1] 李红[1] 郝彦琴[1] 黄丽丽[1] 

机构地区：[1]山西医科大学第一临床医学院感染科,太原030001

出　　处：《山西医科大学学报》2010年第3期277-280,F0003,共5页Journal of Shanxi Medical University

摘　　要：目的探讨体外获取高纯度大鼠骨髓间充质干细胞的方法。方法应用全骨髓贴壁法体外分离培养大鼠骨髓间充质干细胞,进行传代培养,倒置显微镜下观察细胞形态并测定其生长曲线,流式细胞术检测细胞周期,免疫细胞化学法鉴定细胞表面标志CD34、CD44及CD45。结果获取的大鼠骨髓间充质干细胞形态呈均一成纤维细胞样,并呈集落样生长。细胞生长曲线示,在传代后的第4-5天细胞开始明显增殖,进入指数增生期。细胞周期显示81.49%P3代细胞为G0/G1期,经免疫细胞化学染色结果显示CD44阳性,CD34、CD45阴性。结论体外应用全骨髓贴壁法可以分离培养出高纯度的大鼠骨髓间充质干细胞,而且此法简单易行、经济。Objective To explore a better method for obtaining high purity of rat bone mesenchymal stem cells （BMSCs） in vitro. Methods The whole bone marrow adherent culture method was used to culture the BMSCs in vitro. The form of the ceils was observed under inverted microscope, and the cell growth curve was determined. The cell cycle was examined by flow cytometry. Immunocytochem- istry technique was employed to identify the cells. Results The cultured BMSCs showed the typical fibroblast-like morphology and colony. The growth curve showed that the cells multiplied at day 4,5 after the passage. The passage 3 cells in G0/G1 accounted for 81.49%. The BMSCs uniformly expressed CD44, but not CD34 and CD45. Conclusion The whole bone marrow adherent culture method is simple, cheaper and feasible to obtain the adequate high purity of BMSCs.

关 键 词：骨髓间充质干细胞 细胞培养 全骨髓贴壁法 

分 类 号：Q813[生物学—生物工程]