小鼠骨髓间充质干细胞体外扩增的实验研究

The Research of Mouse Bone Marrow-derived Mesenchymal Stem Cells Culture Condition in vitro

目的研究确立全骨髓贴壁法培养小鼠骨髓间充质干细胞体外培养和优化组合最佳实验条件,建立一个简便易行的小鼠骨髓间充质干细胞体外扩增方法.方法观测不同周龄小鼠、不同浓度胎牛血清和不同种植密度条件下的细胞增殖活性.光镜下观察最佳实验条件培养骨髓间充质干细胞的形态.流式细胞检测细胞周期和表面抗原标志.结果 4～6周龄小鼠、5%～10%的胎牛血清和5～8×107/L的细胞种植密度最适宜MSCs的体外培养,细胞形态、表面抗原标志和细胞周期检测均具有骨髓间充质干细胞的特征.结论本实验室优化选择的培养条件可成功分离纯化、扩增小鼠骨髓间充质干细胞,为下一步深入研究奠定基础.

Objective To research the whole bone marrow adherence culure condition of mouse bone marrow-derived mesenchymal stem cells,select the best condition,and establish an expansion method of mouse bone marrow-derived mesenchymal stem cells in vitro.Methods We detected the proliferation ability of the bone marrow mesenchymal stem cells in different ages mouse,different concentration of serum and cell density,and found out the optimum culture condition.BMSCs were cultured in the optimum culture condition,and light microscopy and flow cytometry were used to study the morphology and biological characteristics of BMSCs.Results The optimum culture conditions for BMSCs were 4 ～6 weeks mouse,5% ～10% fetal bovine serum,and 5～8×107/L cell density.The cells cultured in the optimum culture conditions showed the typical morphological and biological characteristics of BMSCs.Conclusion The whole bone marrow adherence method we optimized in our laboratory can isolate,purify and amplify BMSCs of mice in vitro,and may provide basis for further study.