离心分离法与贴壁分离法对培养骨髓间充质干细胞生物学特性影响

Effects of Adherence and Density Gradient Centrifugation Methods on Biological Characteristlcs of Rabbit Marrow Mesenchymal Stem Cells Cultured in Vitro

目的探讨分离高质量和高活性骨髓间充质干细胞的方法。方法选取出生2月龄的健康兔,采用密度梯度离心分离法与贴壁分离法培养骨髓间充质干细胞,检测培养骨髓间充质干细胞生物学特性。结果贴壁分离法培养的原代骨髓间充质干细胞贴壁增殖快,细胞活性高,而密度梯度离心分离法培养的原代骨髓间充质干细胞贴壁增殖慢,细胞活性低,但这些差别在传代后消失。离心分离法得到的细胞纯度较贴壁分离法获得的细胞纯度高。结论与离心分离法相比贴壁法获得的原代骨髓间充质干细胞的细胞活性较高、贴壁增殖快,具有方法简单的优点。

Objective To study the effects of adherence and density gradient centrifugation （DGC） methods on biological characteristics of rabbit marrow mesenchymal stem cells cultured in vitro. Methods Rabbits aged 2 months were used to isolate marrow mesenchymal stem cells （MSCs）. MSCs with adherence and DGC methods were cultured, passaged, amplified and purified in vitro. The living characteristics and adhesive rate of the primary and generations were observed respectively. The growth curve was drawn. The cell cycles and cystoskeleton were tested to study the different MSCs separated methods on the proliferation and metabolism of MSCs. Results MSCs were intact and fibroblast cell-shaped. Adhering spindle-shaped cells with higher cytoactive were observed in the adherence and DGC separation groups. The MSCs of primary culture in the adherence group showed more rapid proliferation,earlier colony confluence and shorter time for passage compared with DGC separation group. Conclusion The MSCs separated by adherence showed a rapid proliferation, earlier colony confluence and shorter time for the first passage.