小鼠骨髓间充质干细胞的分离、培养及鉴定

目的建立一种从小鼠骨髓中分离培养间充质干细胞（MSCs）的高效方法。方法采取贴壁细胞分离法分离和纯化小鼠骨髓间充质干细胞（mMSCs）,检测mMSCs在不同诱导条件下向成骨细胞及脂肪细胞分化能力,用流式细胞术及显微镜分别检测mMSCs纯度和形态特征。结果mMSCs贴壁生长后形态较均一,细胞形态呈成纤维细胞样,流式细胞术检测：CD45、CD11b、CD44及CD29分别为（3.34）%、（2.41）%、（98.46）%及（99.36）%。第4代mMSCs经诱导后可向成骨细胞和脂肪细胞分化。结论通过贴壁培养可以从小鼠骨髓中分离培养出高纯度mMSCs,该方法效率高,稳定性好。

Isolation,Culture and Identification of Mesenchymal Stem Cells from Mouse Bone Marrow

Objective To establish an efficient method for isolation and culture of mesenchymal stem cells （mMSCs） from mouse bone marrow. Methods The mononuclear cells were isolated from mouse bone marrow. The mMSCs were enriched and expanded by using bone marrow adherent culture. The ability of osteogenic and adipogenic differentiation of the mMSCs under different induction conditions was assessed. The phenotype was analyzed by flow cytometry （FCM） and morphology of the mMSCs was examined and analyzed by light microscopy. Result The mMSCs were adherent cells with a similar fibroblastoid spindleshaped morphology during different passages. The expression of CD45, CD11b, CD44 and CD29 was （3.34）%, （2.41）%, （98.46） % , （99.36）% . The mMSCs could be induced to differentiate into osteoblast and adipocytes cells. Conclusion Mouse mesenchymal stem cells ean be isolated and expanded by using bone marrow adherent culture, and it is an efficient and stable method.