论文部分内容阅读
目的探索大鼠骨髓间质干细胞体外诱导分化为胰岛素分泌细胞的方法。为解决胰岛移植物来源匮乏这一问题提供新的思路。方法 采用横向分化技术,将成年大鼠骨髓间质干细胞诱导成为胰岛素分泌细胞。间接免疫荧光法鉴定诱导前后细胞nestin、胰岛素、胰高血糖素、生长抑素及Pdx-1的表达,RT-PCR法检测诱导前后细胞nestin,胰岛素-1,葡萄糖转运子-2,葡萄糖激酶及其转录因子Isl-1,Pdx-1,Pax-4和Pax-6 mRNA的表达;测定24 h胰岛素分泌量和胰岛素刺激实验评价诱导前后细胞的功能。结果 诱导5 h,nestin阳性细胞为(44.6±7.3)％;诱导24 h,nestin阳性细胞增至(61.8±8.4)％;此后,nestin阳性细胞数目开始下降,诱导第14天后,nestin表达基本消失。诱导后的胰岛素分泌细胞可以表达胰岛素、胰高血糖素、生长抑素和Pdx-1等蛋白;表达胰岛素-1、葡萄糖转运子-2、葡萄糖激酶及其多种转录因子mRNA;胰岛素分泌量增加;胰岛素刺激实验反应敏感等。而诱导前MSCc不县备上述特点。结论 大鼠骨髓间质干细胞体外可以诱导成为胰岛素分泌细胞,为胰岛移植开辟新的研究思路。
Objective To explore the method of inducing the differentiation of rat bone marrow mesenchymal stem cells into insulin-secreting cells in vitro. To provide a new way to solve the problem of lack of islet graft. Methods Transverse differentiation was used to induce adult rat bone marrow mesenchymal stem cells into insulin-secreting cells. The expression of nestin, insulin, glucagon, somatostatin and Pdx-1 were detected by indirect immunofluorescence. The expressions of nestin, insulin-1, glucose transporter-2, The transcription factor Isl-1, Pdx-1, Pax-4 and Pax-6 mRNA expression were measured. 24 h insulin secretion and insulin stimulation assay were used to evaluate the function of cells before and after induction. Results Nestin positive cells were (44.6 ± 7.3)% at 5 h after induction. The number of nestin positive cells increased to (61.8 ± 8.4)% at 24 h after induction. After that, the number of nestin positive cells began to decrease. After the 14th day, the expression of nestin almost disappeared . Insulin-secreting cells after induction can express insulin, glucagon, somatostatin and Pdx-1 and other proteins; express insulin-1, glucose transporter-2, glucokinase and various transcription factor mRNAs; increase insulin secretion ; Insulin stimulation experiment sensitive response. Before the induction of MSCc County does not have the above characteristics. Conclusion Rat bone marrow mesenchymal stem cells can be induced to become insulin-secreting cells in vitro, which opens up new research ideas for islet transplantation.