探讨大鼠骨骼肌卫星细胞（MDSCs）定向诱导分化为胰岛素生成细胞（IPCs），为1型糖尿病的干细胞治疗提供一种新的研究思路。

通过二次酶消化法和差速贴壁培养法分离、培养大鼠MDSCs，利用不同的诱导培养液使MDSCs定向分化为IPCs，并对诱导后细胞进行形态观察，通过双硫腙染色和免疫组化染色对MDSCs-IPCs形态进行鉴定，采用Q-PCR和Western Blot方法检测MDSCs-IPCs中C-peptide和Insulin的表达，通过胰岛素释放实验检测MDSCs-IPCs的生物学功能，β细胞和MDSCs-IPCs两组间比较采用t检验。

MDSCs在接种4 h后开始贴壁部分细胞伸出小的突起，48 h后绝大多数细胞贴壁呈梭形、胞浆丰富、折光度高。随着培养时间的延长，细胞的梭形形状更为明显且生长迅速。免疫组化结果显示细胞表达Desmin、α-Sarcomeric Actinin、MyoD1、Myf5和PAX7。成胰诱导后MDSCs形成胰岛样的圆形细胞团，双硫腙染色呈猩红色，Insulin免疫组化染色阳性。Q-PCR结果显示MDSCs-IPCs中C-peptide和Insulin mRNA表达量分别是β细胞的0.73倍（P > 0.05）和0.79倍（P > 0.05）。胰岛素释放实验显示，5.6 mmol/L和16.7 nmlol/L葡萄糖刺激培养2 h后，β细胞和MDSCs-IPCs分泌胰岛素量分别为[（20.3±4.2）mU/L]、[（16.1±3.7）mU/L]、[（60.5±9.3）mU/L]和[（40.9±7.3）mU/L]，葡萄糖可调节MDSCs-IPCs胰岛素的分泌。

MDSCs易于分离培养、增殖能力强，体外可诱导分化为有功能的IPCs，适合作为再生医学的种子细胞。

To investigate the directional differentiation of Muscle-derived satellite cells (MDSCs) into insulin producing cells (IPCs), and provide experimental and theoretical basis for the future use of adult stem cells in the treatment of diabetes.

Rat MDSCs were separated, cultured by twice enzyme digestion method and differential adherent culture method and differentiated into IPCs by different induced medium. The morphology of MDSCs-IPCs was observed by Dithizone staining and immunohistochemistry and the gene expression was detected by Q-PCR and Western Blot. The biological function of MDSCs-IPCs was detected by the insulin release test.

The rat MDSCs were getting adhered after 4 hours, and some cells developed small processes. The vast majority of cells transferred into spindle-shaped mononuclear cells after 48 h, with abundant cytoplasm and high cell refractive index. The fusiform shape became more apparent as the incubation time increasing, with strong proliferation and rapid growth ability. Desmin, α-Sarcomeric Actinin, MyoD1, Myf5and PAX7 were positive in MDSCs. After pancreatic induction, MDSCs formed an islet like a circular cell group. Dithizone staining was brown red. Compared with β cells, the expression of C-peptide and insulin was also detected in MDSCs-IPCs. Glucose regulated the secretion of insulin in MDSCs-IPCs.

The results suggest that it was easy to isolate, culture and amplify rat MDSCs with a wide range of material source. It can be induced to functional IPCs in vitro and is suitable for the seed cells of regenerative medicine.