全骨髓贴壁法分离培养大鼠骨髓间充质干细胞的生物学特性及优势

背景：骨髓来源的间充质干细胞含量极低，体外纯化、扩增活性好、分化潜能高的骨髓间充质干细胞，对进一步研究至关重要。目的：进一步验证全骨髓贴壁法体外分离、培养、纯化骨髓间充质干细胞的生物学特性、表型及多向分化潜能。方法：通过全骨髓贴壁法体外分离、培养、纯化大鼠骨髓间充质干细胞，进行形态学观察，流式细胞仪检测细胞标记物表达，分别进行成骨、成脂诱导分化。结果与结论：成功纯化、扩增了高细胞活性、高分化潜能的骨髓问充质干细胞。所得细胞呈成纤维细胞样；表达CD29、CD90，不表达CD45；成骨、成脂诱导分化后，茜素红染色、油红O染色阳性。证实全骨髓贴壁法操作简单、对细胞活性损伤小，可以得到高纯度、高活性、高分化潜能、生物学形态和特征稳定的骨髓间充质干细胞。

Biological characteristics and superiority of rat bone marrow mesenchymal stem cells isolated and cultured using whole bone marrow adherence method

BACKGROUND：Bone marrow mesenchymal stem cells are rare in vivo. It is important to purify, proliferate and differentiate bone marrow mesenchymal stem cells in vitro for further research. OBJECTIVE：To evaluate the biological characteristics, phenotype and multiple differentiation potential cultivation of bone marrow mesenchymal stem cells that are isolated, cultured and purified using the whole bone marrow adherence method. 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 cellsurface markers were performed. Osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was induced. RESULTS AND CONCLUSION：We successful y purified and proliferated bone marrow mesenchymal stem cells with high cellviability and differentiation ability. Fibroblast-like cells were harvested, expressing CD29 and CD90, but not CD45. Fol owing osteogenic and adipogenic induction, cells were positive for oil red O staining and alizarin red staining. The whole bone marrow adherence method is easy to operate, has little impact on cellviability, and can be used to harvest high-purification bone marrow mesenchymal stem cells with high cellviability and differentiation ability.