密度梯度离心法结合贴壁筛选法分离培养大鼠骨髓间充质干细胞的生物学特性

摘要

目的：通过密度梯度离心法结合全骨髓贴壁法体外分离、培养、纯化骨髓间充质干细胞。方法采用密度梯度离心法结合全骨髓贴壁法，对大鼠骨髓间充质干细胞进行体外分离后传代培养，倒置相差显微镜下观察形态变化，流式细胞仪检测细胞标记物表达。结果 P3代大鼠BMSCs 细胞培养5天，CD29、CD90、CD34阳性率分别为94.27%、91.16%、1.72%。结论密度梯度离心法结合全骨髓贴壁法成功分离培养了高纯度、高活性、生物学形态和特征稳定的骨髓间充质干细胞。%Objective To evaluate the biological characteristics, phenotype of bone marrow mesenchymaI stem cells that are isolated. Methods Bone marrow mesenchymal stem cells were isolated, purified and cultured by the whole bone marrow adherence method. Morphological observation and flow cytometry determination of celI surface markers were performed., Results The positive rates of BMSCs, CD34 and CD29 were 5, 91.16% and 94.27% in P3 cells cultured in CD90 generation. Conclusion The whole bone marrow adherence method is easy to operate, and can be used to harvest high-purification bone marrow mesenchymal stem cells with high cell viability and diferentiation ability.