大鼠肌源性干细胞体外转分化为胰岛素分泌细胞

Transdifferentiation of rat muscle derived stem cells into insulin-secreting cells in vitro

目的探讨体外自体肌源性干细胞（MDSCs）定向诱导分化为胰岛素分泌细胞团的可行性。方法用差速贴壁的方法对SD大鼠MDSCs进行分离和培养,加入联合诱导剂培养7-12 d。通过形态学观察、双硫腙（DTZ）染色、免疫细胞化学染色、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。结果联合诱导后12 d有典型的胰岛样结构出现;诱导后第12天细胞团表达胰岛素,而未诱导组细胞不表达;诱导后第7天检测到胰岛素（INS）、胰高血糖素（GLU）和生长抑素（SS）基因的表达。结论体外诱导大鼠肌源性干细胞可定向分化为胰岛素分泌细胞存在可行性,本实验将可能为临床糖尿病干细胞治疗提供新的途径。

Objective To explore the potential diferentiation of insulin-secreting cells from muscle derived stem cells. Methods SD rat muscle derived stem cells （MDSCs） were isolated and then induced for 7 - 12 days with combination inductor. Morphological changes of the induced cells were observed under an inveted microscope. DTZ staining,immunocytochemical staining were conducted to examine the production of insulin. Glucose challenge and radio immunoassay were perfomed to evaluate the function of the aggregates, and RT-PCR was conducted to detect the expression of endocrine gene. Results Typical insulin-producing cells were observed at the end of the protocol （12 days）. Ditizone staining and immunohistochemistry for insulin were both positive. Insulin was secreted by these cells in response to different concentrations of glucose stimulation （ P 〈 0. 05 ）. RT-PCR showed the insulin, glucagon and SS gene expression on the 7th day after the inducement of combination inductor. Conclusion Insulin-secreting cells can be transdiferentiated from muscle derived stem cells, which may be a potential approach for clinical treatment of diabetes.