论文部分内容阅读
目的研究犬骨髓间充质干细胞(MSC)体外诱导分化为胰岛样细胞的可能性。方法分离纯化犬骨髓MSC,予EGF、bFGF和β细胞调节素、尼克酰胺、B27添加剂诱导。双硫腙染色、Westernblot、免疫荧光染色、胰岛素分泌量测定和葡萄糖刺激胰岛素释放试验鉴定诱导前后的细胞功能。结果未诱导细胞呈长梭形贴壁生长,诱导第2周细胞逐渐变圆,聚集成团,双硫腙染色呈棕红色;Westernblot检测诱导后细胞表达PDX-1;免疫荧光染色显示诱导后细胞表达胰岛素、胰升血糖素、生长抑素;ELISA结果表明诱导后细胞可分泌胰岛素,体外葡萄糖刺激胰岛素释放试验阳性。结论犬骨髓MSC在体外能被诱导分化为胰岛样细胞。
Objective To investigate the possibility of in vitro differentiation of canine bone marrow mesenchymal stem cells (MSCs) into islet-like cells. Methods Canine bone marrow MSCs were isolated and purified, and induced by EGF, bFGF, β-cell regulators, nicotinamide and B27 additives. Dithizone staining, Western blotting, immunofluorescence staining, insulin secretion assay and glucose-stimulated insulin release assay were used to identify the cell function before and after induction. The results showed that the cells did not induce long fusiform adherent growth, the cells became round at the second week of induction, aggregated into clusters, dithizone staining was brown-red; PDX-1 expression was detected by Western blot; the immunofluorescence staining showed that the cells The expression of insulin, glucagon and somatostatin were measured. The results of ELISA showed that the cells could secrete insulin after induction, and the glucose-stimulated insulin release test was positive in vitro. Conclusion Canine bone marrow MSCs can be differentiated into islet-like cells in vitro.