体外诱导牛BMSC向上皮样细胞分化的研究

Study of differentiation of bone marrow mesenchymal stem cells into epithelial-like cells in vitro

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 genetically modified beef cattle and enrich multi-differentiation potential of the bovine BMSC, immunofluorescence staining and molecular biology technology were used to explore the possibility of BMSC differentiating into epithelial-like cells in vitro in the presence of the epidermal growth factor （EGF） and insulin. Purified and stable P4 generation bovine BMSC were utilized to induce in vitro on medium with EFG and insulin. We used immunofluorescence staining of cytokeratin 18 （CK18） and RT-PCR of cytoker- atin 19 （CK19） to observe and identify the cells after inducing. The results clearly showed fluorescence by CK18 staining. RT-PCR results demonstrated that cells after induction clearly expressed CK19 gene. Consequently, in vitro BMSC can differentiate into epithelial-like cells by using multi-factor.