兔骨髓间充质干细胞体外分离培养及生长特性的初步研究

Study of the isolation,culture and growth characteristics of rabbit bone marrow-derived mesenchymal stem cells in vitro

目的:探索兔骨髓间充质干细胞(MSC)体外分离及培养扩增的方法。方法:骨髓穿刺法抽取雄性新西兰白兔髂骨骨髓5ml,联合应用密度梯度离心及贴壁培养法分离纯化MSC并进行体外扩增,倒置显微镜下观察原代及传代细胞的形态、生长情况,计数细胞数目,绘制细胞生长曲线。HE染色光镜观察细胞形态。结果:成功建立了兔MSC体外分离及培养扩增的方法,生长动力学分析发现,传代细胞随传代次数的增加,增殖能力逐渐下降。结论:兔MSC能在体外成功培养、扩增,可作为组织工程的种子细胞。

Objective： To explore an effective method to isolate and culture rabbit bone marrow-derived mesenchymal stem cells （MSC）, Methods： 5ml bone marrow was take from iliums of New Zealand white rabbit： then after the density gradient centrifugation, mononuclear cells were got and cultured. MSC with relative purification was achieved by changing culture fluid repeatedly and cultured continuously. The growth and morphological changes of the cells were observed under invert microscope every day. Growth curves of cells were drawn. Morphological characteristics were observed by HE staining. Results： The method of culture and expansion of MSC in vitro were e：tablished. The proliferation ability of MSC declined with time of the culture. Conclusion： MSC can be cultivated successfully in vitro and can be used in tissue engineering.