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目的建立一种简单实用的小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)体外分离培养方法,为BMSCs的进一步研究和应用提供实验材料和依据。方法采用直接贴壁法和分阶段更换培养基的方法分离纯化BMSCs,显微镜下观察其形态,应用流式细胞技术鉴定细胞表面标记。结果显微镜下BMSCs贴壁生长后呈梭形,形态均一,经流式细胞仪检测第2代BMSCs高表达CD34、Sca-1,不表达CD45。结论本实验方法能简单高效的获得高纯度BMSCs,且细胞生物学特性稳定,适用于对BMSCs的进一步研究和应用。
Objective To establish a simple and practical method for isolation and culture of mouse bone marrow mesenchymal stem cells (BMSCs) in vitro and to provide experimental materials and basis for further research and application of BMSCs. Methods BMSCs were isolated and purified by direct adherent method and medium replacement by stages. The morphology of the cells was observed under a microscope. Cell surface markers were identified by flow cytometry. Results The BMSCs were spindle-shaped and their morphology was uniform under the microscope. The second-generation BMSCs were highly expressed in CD34 and Sca-1 but not in CD45 by flow cytometry. Conclusion The experimental method can obtain high-purity BMSCs easily and efficiently with stable cell biological characteristics, which is suitable for further research and application of BMSCs.