摘要
目的:探讨大鼠胰腺提取液(RPE)对大鼠肌源性干细胞(MDSCs)定向诱导分化为胰岛素分泌细胞的作用。方法:RPE诱导MDSCs通过形态学观察、双硫腙(DTZ)染色、免疫细胞化学显色、葡萄糖刺激实验和RT-PCR,对诱导细胞进行体外结构和功能鉴定。结果:RPE诱导后第5日有胰岛样结构出现;免疫细胞化学显示,诱导后第12日细胞团表达胰岛素、胰高血糖素,而未诱导组细胞不表达上述蛋白;RT-PCR结果表明,诱导后第6日检测到PDX-1的表达,诱导后第12日检测到胰岛素1、胰岛素2、胰高血糖素和葡萄糖转运体2基因的表达。结论:体外经RPE诱导,大鼠肌源性干细胞可定向分化为胰岛素分泌细胞。
Objective： To explore the possibility in differentiating functional insulin-secreting cells from muscle-derived stem cells induced by rat pancreatic extract （RPE）. Methods： Adult SD rat muscle-derived stem cells （MDSCs） were isolated, cultured and then induced with serum free DMEM media supplemented with RPE for 6 days. The cells were then cultured in DMEM supplemented with 10% FBS for 5-8 additional days. Morphological changes of the induced cells were observed. DTZ and immunoeytochemical staining were used to examine the production of insulin and glucagons. Glucose challenge test and radioimmunoassay were performed to evaluate the function of the aggregates, and RT-PCR was conducted to detect the expression of endocrine gene. Results： While MDSCs cells were typically fibrocyte-like, grape-like coherent clustered cells were observed in the experiment group on the 6th day after RPE induction, which were similar to pancreatic islets. DTZ staining of the cell aggregates was positive as the natural pancreatic islet. Immunocytochemistry also confirmed that these aggregates were positive for insulin on the 12th day after the RPE induction; while the control group were negatively stained for the characteristics of endocrine. RTPCR showed that the insuln 1, insulin 2, glucagon and glucose transporter 2 gene expressed on the 12th day after the RPE induction. Conclusion： Insulin-secreting cells can be transdifferentiated from muscle derived stem cells by the inducement of RPE.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2008年第5期629-632,共4页
Chinese Journal of Anatomy
基金
辽宁省教育厅基金(20040131
20060519)
关键词
肌源性干细胞
分化
大鼠胰腺提取液
胰岛素
muscle-derived stem cells
differentiation
rat pancreatic extract
insulin
