大鼠骨髓间充质干细胞体外两种分离方法和培养条件下生物学特点的比较

目的：建立并探讨一种体外分离、培养和扩增骨髓间质干细胞的优化方法。方法：采用密度梯度离心法和全骨髓贴壁法分离培养大鼠骨髓干细胞，应用倒置显微镜和免疫组化法观察和鉴定骨髓间质干细胞。结果：大鼠骨髓干细胞呈均一的梭形成纤维细胞样，形成集落样生长，显微结构表现出干细胞特征；免疫组化显示CD14、CD45阴性，CD44、CD90阳性。与密度梯度离心法比较，贴壁法获得的骨髓干细胞活性高，增殖力强，克隆形成早，传代时间短。结论：全骨髓贴壁法和密度梯度离心法均可获得高纯度贴壁生长的骨髓干细胞，其中全骨髓贴壁法方法简单易行，骨髓于细胞增殖快，活性好，传代力持久，是一种有实用价值的骨髓干细胞培养方法。

Comparison of the Biology Characteristics of Rat Bone Marrow Mesenchymal Stem Cells under Different Isolation and Culture Methods

Objective：To investigate the more suitable methods for isolation, culture and proliferation of bone marrow mesenchymal stem cells（BMSCs）, so as to acquire high quality and activity BMSCs. Methods：BMSCs were isolated from rat bone marrow by density gradient centrifugation and adherence separation methods respectively. The characteristic and the morphology of BMSCs were observed with inverted microscope every day. The surface markers of BMSCs were identified by immunocytochemical technique. Results： After purification and proliferation, the primary cultured BMSCs were in uniformly long spindle-shapped form and formed cell colony,which showed the characteristics of stem cell. Immunocytochemistry showed BMSCs were positive for CD44 and CD90, while negative for CD14 and CD45. The primary cells cultured by adherence separation method showed higher cytoaetive, more rapid proliferation, earlier colony confluence and shorter time for passage than that cultured by density gradient centrifugation method. Conclusion： BMSCs were obtained both with density gradient centrifugation and adherence separation methods. The adherence separation method is a more valuable technique for the isolation and purification of BMSCs.