人骨髓基质细胞的培养及鉴定

Cultivation and identification of human mesenchymal stem cells

目的:探讨骨髓基质细胞的分离、体外培养方法,为基因治疗提供载体细胞。方法:采用密度梯度离心法结合贴壁筛选法分离人骨髓进行体外培养扩增,用倒置显微镜观察细胞形态学特征并用流式细胞仪鉴定其CD90表达。结果:分离培养的骨髓基质细胞似成纤维状,原代细胞呈集落生长,并可表达CD90,传代后细胞形态较一致,且随着传代次数的增加CD90表达越高,第3代可达94.31%。结论:骨髓基质细胞可通过体外培养纯化获得,随着传代次数增加,纯度越高,该方法是一种较理想的骨髓基质细胞培养方法。

Objective：To explore a practical method of separation and in vitro cultivation of mesenchymal stem cells （MSCs） from human bone marrow, and to offer carrier cell for gene therapy. Method： MSCs from human bone marrow were separated and purified by means of gradient centrifugation and adherence to the culture plastic, which were detected by Flow cytometry with cell surface markers. Result.. MSCs had spindle shape and had a typi- cal fibroblast-like morphology. MSCs expressed positive for CD90. The cells were noted to have a simple shape af- ter subculture, which had a higher expression of CD90. 94.31 % of the third generation cells showed positivity to CD90. Conclusion：MSCs were obtained from human bone marrow in vitro, which had a high purity with passage culture. This methods is suitable for MSCs.