成年大鼠骨髓间充质干细胞的体外分离  

Comparative study on different culture methods to get stable adult bone mesenchymal stem cells rapidly

作　　者：周丽娜[1] 谢华[1] 

机构地区：[1]广东医学院人体解剖学教研室,广东东莞523808

出　　处：《解剖学研究》2009年第6期420-423,共4页Anatomy Research

摘　　要：目的应用全骨髓贴壁法和密度梯度离心法以不同培养条件,探讨体外获得高质量、高活性的成年大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)的方法。方法取成年雄性SD大鼠双侧股骨,应用全骨髓贴壁法,分为10%国产TBD胎牛血清+DMEM组、10%国产TBD胎牛血清+DMEM/F12组、12.5%国产TBD胎牛血清+DMEM/F12组、10%进口以色列胎牛血清+DMEM/F12组4组,3只/组;应用密度梯度离心培养法,以10%进口以色列胎牛血清+DMEM/F12作为培养基,取3只大鼠作为一组。比较不同培养方法及培养条件对BMSCs生长增殖的影响。结果全骨髓贴壁法中10%进口以色列胎牛血清+DMEM/F12组原代细胞即可7～9 d达融合,传代到第4代细胞活力仍很好。其余3组原代细胞达融合的时间均超过10 d,且传代后细胞活力差,容易出现老化。密度梯度离心培养法原代贴壁细胞太少,无法达融合。结论本实验采用的培养方法中,10%进口以色列胎牛血清+DMEM/F12培养基的全骨髓贴壁分离法,是体外获取成年大鼠BMSCs最佳方法。Objective To seek a stable, rapid and effective method in order to obtain large number of adult rat Bone Mesenchymal stem ceils （BMSCs） in vitro. Methods Two method were used in the experiment to separated BMSCs from rat bone marrow. 12 SPF male SD rats were divived into groups in adherence separation： 10% domestic FBS DMEM group , 10% domestic FBS DMEM/F12 group,12.5% domestic FBS DMEM/F12 group,10% import FBS DMEM/F12 group, 3 rats in each group;3 SPF male SD rats were used in gradient centrifugation with 10% import FBS DMEM/F12. Results The confluence of primary cells was happened within 7-9 days in 10% import FBS DMEM/F12 group of adherence separation. The confluence time of primary ceUs were over the 10th day in other three groups. Only 3-5 primary ceils in every 100 field were adherent in gradient centrifugation with 10% import FBS DMEM/F12, confluence is difficult＇. Conclusion The 10% import FBS DMEM/F12 group in adherence separation was the most stable and effective method to obtain large number of adult Bone Mesenchymal stem cells in vitro rapidly in above methods.

关 键 词：骨髓间充质干细胞 细胞培养 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]