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目的探讨葡萄糖转运蛋白2(Glut2)在人脐带间充质干细胞(h UMSCs)向胰岛前体细胞分化过程中的表达变化。方法分离、培养、鉴定及诱导h UMSCs,分别收集诱导过程中第7天、14天和21天细胞和细胞上清液,采用免疫细胞化学、ELISA、免疫荧光、Western blotting及Real-time PCR检测诱导后细胞相关蛋白和基因的表达。结果免疫组织化学检测诱导后细胞胰腺十二指肠同源盒基因-1(PDX-1呈阳性表达;免疫荧光检测诱导后细胞Ngn3和胰岛素呈阳性表达;Western blotting检测Glut2在诱导过程中逐渐升高,诱导14 d达到峰值,与正常组比较P<0.01;Real-time PCR显示,Glut2基因自第7天即增高(P<0.05)。结论 h UMSCs经诱导后分化为胰岛前体细胞,表达Glut2后能引起胰岛素分泌,已初步具有胰岛B细胞功能。
Objective To investigate the expression changes of Glut2 during the differentiation of human umbilical cord mesenchymal stem cells (h UMSCs) into islet precursors. Methods Human umbilical cord blood mononuclear cells (UUMSCs) were isolated, cultured, identified and induced. The supernatants of cells and supernatant were collected on day 7, day 14 and day 21 respectively. Immunocytochemistry, ELISA, immunofluorescence, Western blotting and Real-time PCR Induction of cell-associated protein and gene expression. Results The expression of PDX-1 in the pancreatic duodenal homeobox was detected by immunohistochemistry. The expression of Ngn3 and insulin was detected by immunofluorescence. The expression of Glut2 was detected by Western blotting (P <0.05) .Conclusion h UMSCs differentiate into islet precursor cells after induced by Real-time PCR, the expression of Glut2 gene increases from the 7th day Glut2 can cause insulin secretion, has been initially has islet B cell function.