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为培育转基因肉牛提供种子细胞以及进一步丰富牛骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)的多向分化潜能,利用细胞免疫荧光染色和分子生物学方法,初步探讨表皮生长因子和胰岛素体外诱导牛BMSC向上皮样细胞分化的可能性。利用含细胞因子的诱导液对纯化稳定的P4代牛BMSC进行体外诱导,并对诱导后的细胞进行细胞角蛋白18的细胞免疫荧光观察和细胞角蛋白19的RT-PCR鉴定。结果表明,诱导后细胞经细胞角蛋白18免疫荧光染色后出现明显的荧光。RT-PCR结果显示诱导分化后细胞角蛋白19基因在细胞中表达。因此,在体外,表皮生长因子和胰岛素可诱导牛BMSC初步分化为上皮样细胞。
To provide seed cells for breeding of transgenic beef cattle and to further enrich the potential of multidrug differentiation of bovine bone marrow mesenchymal stem cells (BMSCs), the effects of epidermal growth factor and insulin in vitro were investigated by immunofluorescent staining and molecular biology methods Induction of bovine BMSC into epithelial-like cells. Purified Staphylococcus aureus BMSC was induced in vitro using cytokine-containing inducer, and the induced cells were observed by immunofluorescence and cytokeratin 19 RT-PCR. The results showed that after induction of cells by cytokeratin 18 immunofluorescence staining obvious fluorescence. The results of RT-PCR showed that the cytokeratin 19 gene was expressed in the cells after induced differentiation. Therefore, in vitro, epidermal growth factor and insulin induce initial differentiation of bovine BMSC into epithelial-like cells.