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目的在乳鼠骨髓间充质干细胞(MSC)定向诱导成产胰岛素细胞过程中,检测胰十二指肠同源性盒因子1(PDX-1)和nestin的表达。方法通过细胞培养方法分离培养MSC,加入10μg/L人表皮生长因子(EGF)、10μg/L碱性成纤维细胞生长因子(b FGF)、10μg/L肝细胞生长因子(HGF)、10μg/L人B细胞调节素、20 mmol/L烟酰胺和20 g/L B27进行诱导。诱导后,采用反转录PCR检测胰岛素、PDX-1和nestin的mRNA的表达,并以免疫荧光细胞化学染色检测胰岛素、PDX-1和nestin蛋白表达。结果在乳鼠MSC分化为产胰岛素细胞过程中,胰岛素和PDX-1 mRNA的表达上调,nestin mRNA表达下降;nestin、PDX-1和胰岛素蛋白能在产胰岛素细胞中共表达。结论 MSC可以定向诱导而表达胰岛素。
Objective To detect the expression of PDX-1 and nestin during the induction of insulin-producing cells by bone marrow-derived mesenchymal stem cells (MSC) in suckling mice. Methods MSC were isolated and cultured by cell culture method. 10 μg / L human epidermal growth factor (EGF), 10 μg / L bFGF, 10 μg / L hepatocyte growth factor (HGF) Human B-cell regulators, 20 mmol / L nicotinamide and 20 g / L B27. After induction, the mRNA expression of insulin, PDX-1 and nestin was detected by reverse transcription PCR. The expressions of insulin, PDX-1 and nestin were detected by immunofluorescence cytochemistry. Results The expression of insulin and PDX-1 mRNA was up-regulated and the expression of nestin mRNA was decreased during the differentiation of MSC into insulin-producing cells. Nestin, PDX-1 and insulin protein could be co-expressed in insulin-producing cells. Conclusion MSC can be induced to express insulin.