Induced differentiation of rat skeletal muscle satellite cells into insulin producing cells

Abstract

Objective 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. Methods 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. Results 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. Conclusion 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. Key words: Muscle-derived satellite cells; Insulin producing cells; Differentiation; Function