骨髓基质干细胞的分离纯化及培养

Preparation and culture of murine bone marrow stromal stem cdlls

目的 建立骨髓基质干细胞(MSCs)良好的分离纯化和培养方法。方法 将小鼠骨髓基质干细胞自股骨中分离,应用贴壁选择法结合细胞克隆挑选法进行分离纯化,应用细胞生长因子(EGF和PDGF-BB)刺激法进行MSCs的体外培养和传代,倒置显微镜下观察分离培养的细胞并照像记录。结果,培养获得了纯化的呈梭形成纤维样细胞的骨髓基质干细胞。在生长因子EGF和PDGF-BB的共同作用下,传代MSCs生长旺盛,形态均一。结论 该方法是简便高效的骨髓基质干细胞的分离纯化和培养方法。

A method for isolating and culturing mouse bone marrow_detived mesenchymal stem cells (mMSCs) were established.MSCs were isolated from mouse thighbone marrow and cultured in Dulbecco's modified eagle's medium with low glucose and purified by wall_sticking and colonies collection,and were proliferated through subculturing by adding epidermal growth factor (EGF)and pletelet_derived growth factor(PDGF-BB).Cellular proliferation were observed and recorded under the inverted microscope.Purified MSCs were acquired and became fibroblast_like.EGF and PDGF_BB were effective agents for mMSCs proliferation in vitro.