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目的建立可行的人骨髓间充质干细胞(human marrow mesenchymal stem cells,h MSCs)的培养方法,并对其生物学特性进行初步研究。方法于2013年1月—2014年6月采用密度梯度离心法和贴壁法分离培养h MSCs,用免疫组织化学(免疫组化)法鉴定表面标志并分析其核型,光镜观察其生长和形态变化。结果本实验分离的h MSCs纯度高、贴壁好,以长梭形为主,至少在传代培养6代内增殖快,性状稳定。免疫组化染色显示h MSCs表达CD29和CD44,不表达CD45及CD34,核型正常。结论密度梯度离心法可获得增殖活跃且纯度较高的h MSCs,有特征性表型,适合体外较长期的培养h MSCs。
Objective To establish a feasible method of culturing human marrow mesenchymal stem cells (hMSCs) and to study its biological characteristics. Methods From January 2013 to June 2014, h MSCs were isolated and cultured by density gradient centrifugation and adherent method. The surface markers were identified by immunohistochemistry (IHC) and their karyotypes were analyzed. The growth and Morphological changes. Results hMSCs isolated in this experiment had high purity, good adherence and long fusiform shape, and they were proliferated rapidly and had stable characters at least in passage 6 subculture. Immunohistochemical staining showed that h MSCs express CD29 and CD44, but do not express CD45 and CD34, with normal karyotype. Conclusion Density gradient centrifugation can obtain hMMSCs with active proliferation and high purity, and has characteristic phenotype, which is suitable for hMSCs cultured in vitro for a longer period.