Differentiation of bone marrow mesenchymal stem cells into insulin-producing cells via transfection of Pdx-1, Ngn3, and MafA

Abstract

Objective To investigate the effect of pancreatic and duodenal homeobox1(Pdx-1), neurogenin 3(Ngn3), and v-maf muscu-loaponeurotic fibrosarcoma oncogene homolog A(MafA) on the differentiation of bone marrow mesenchymal stem cells(BMSC) into insulin-producing cells(IPCs). Methods BMSCs were separated and purified by attachment culture method in vitro, and then they were divided into 4 groups(A: un-infection group; B: MafA infection group; C: Pdx-1-Ngn3 infection group; D: co-infection group). BMSCs were cultured for 7 days under high glucose conditions. The infection efficiency was observed through fluorescence microscope. The protein expression of Pdx-1, Ngn3, and MafA were detected by Western blot. mRNA expression of Insulin, Pdx-1, glucokinase(GK), nestin, and glucagon were detected by RT- PCR. The secretion of insulin were measured by ELISA. Results The induced effect was pronounced after infected for 7 days. The morphology of most cells appeared as grape-like aggregation and clustered islet-like cells by the end of induction, and they were confirmed by dithizone(DTZ) staining. The insulin secretion levels of each infection group were gradually improved before and after infected 3rd, 5th, 7th, and 9th day, and in D group, the increment was most significant(P<0.05). mRNA expression levels of insulin, Pdx-1, GK, and glucagon in the infection groups were significantly higher than A group(P<0.05). Conclusions BMSCs are able to be induced into IPCs by infection of Pdx-1, Ngn3, and MafA. (Chin J Endocrinol Metab, 2015, 31: 891-896) Key words: Pancreatic and duodenal homeobox 1; Neurogenin 3; v-maf muscu- loaponeurotic fibrosarcoma oncogene homolog A; Bone marrow mesenchymal stem cells; Insulin-producing cells