SD大鼠骨髓基质细胞的分离培养和鉴定

目的观察SD大鼠骨髓基质细胞的体外分离培养情况并加以鉴定。方法采用密度梯度离心(Percoll法)和贴壁培养结合的方法分离培养SD大鼠BMSCs;采用倒置相差显微镜观察BMSCs形态学特征;流式细胞仪检测细胞表面标志抗原CD45和CD90表达阳性率。结果第3代以后的细胞形态较一致,呈大而扁的梭形为主,第3～5代细胞增殖力保持旺盛;第3代细胞表面标志抗原CD45和CD90表达阳性率分别为:0.2%和94.7%。结论密度梯度离心结合贴壁培养的方法可获得较高纯度的BMSCs,对其进行研究和应用应选用第3～5代的BMSCs。

ISOLATION, CULTIVATION AND IDENTIFICATION OF BONE MARROW STROMAL CELLS OF SD RATS

Objective To investigate the isolation and cultivation of bone marrow stromal cells of SD Rats, and to identify them. Methods MSCs were isolated and cultivated from the bone marrow of SD rats by Percoll gradient centrifugation and investigated their adherence - dependent growth characters. Morphological character of BMSCs was observed with inverted phase contrast microscope, The expression of CD45 and CD90 of the cells were analyzed by flow cytometry. Results The morphous of BMSCs of ulterior the third generation were comparatively coincident, they almost show large and flat fusiform shape. The BMSCs of the 3rd ,4th and 5th generation kept their proliferative ability highly. The positive rates of CD45 and CD90 in BMSCs at the 3rd generation were 0. 2% and 94.7% respectively. Conclusion Purer BMSCs can be isolated and cultivated by Percoll gradient centrifugation and keep its adherence - dependent growth character. BMSCs of the 3rd,4th and 5th generation are well for research and application.