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目的探讨SD大鼠骨髓间充质干细胞(BMSCs)体外培养、纯化及鉴定的方法。方法对1 w龄SD大鼠脱颈处死后,剥离大鼠两侧后肢股骨和胫骨提取BMSCs,采用全骨髓贴壁法分离、培养、纯化BMSCs,观察细胞形态变化,绘制生长曲线,并通过成骨、成脂诱导、克隆形成率等方法对其进行鉴定。结果应用全骨髓贴壁培养法体外分离、纯化、培养的SD大鼠BMSCs生长迅速、生长曲线较好,经过成骨、成脂、克隆形成率等方法鉴定证实其为SD大鼠BMSCs。结论全骨髓细胞贴壁培养法是分离、纯化与培养BMSCs的一种有效方法,能较快获得增殖能力较强的原代细胞用于实验研究。
Objective To investigate the method of culture, purification and identification of SD rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods BMSCs were extracted from the femur and tibia of hind limbs on both sides of rats after 1 week old SD rats were sacrificed. The BMSCs were isolated, cultured and purified by whole bone marrow adherent method. The morphological changes were observed and the growth curves were drawn. Bone, adipogenesis, clonality and other methods to identify its. Results The BMSCs isolated, purified and cultured in vitro from whole bone marrow adherent culture grew rapidly and had good growth curve. BMSCs were identified as osteogenic, adipogenic and clonogenic rates in SD rats. Conclusion Whole bone marrow adherent culture method is an effective method to isolate, purify and culture BMSCs, which can quickly obtain primary cells with strong proliferative ability for experimental study.