A simple method for the generation of insulin producing cells from bone marrow mesenchymal stem cells

摘要: To produce insulin-producing cells (IPCs) from bone marrow mesenchymal stem (BM-MSCs) using a simple and cost effective method. During the initial 7 days of three-dimensional (3D) culture, BM-MSCs were cultured on 1% agar or agarose to form multicellular spheroids. Spheroids spheroid-derived single (SS SSC, respectively) in absence any proteinaceous growth factor specific medium for further 7 d. The insulin content differentiated was evaluated at mRNA protein levels. Furthermore, expression pancreatic beta cells-related genes other than INS as well vitro responses IPCs different glucose concentrations investigated. Cellular clusters generated SS conditions (agar+SS-IPCs) stained better with cell stains more reactive serum-containing antibodies compared agarose-SS-IPCs. Gene analysis revealed that comparison agarose + SS-IPCs, agar+SS-IPCs expressed significantly higher levels INS-1, INS-2, PDX-1, NKX6.1, XBP-1. Of interest, 2215.3 ± 120.8-fold INS-1 gene BM-MSCs. β-cell associated also agar+SSC-IPCs. Moreover, agar+SSC-IPCs after culture media high concentration glucose. Compared most expensive time-consuming protocols, 3D MSCs followed by 2D cellular minimally supplemented produced highly potent which may pay way treatment diabetic patients.