Evaluation of pdx-1 gene expression in insulin producing cells, derived from embryonal carcinoma stem cells

Understanding β-cell function at the molecular level will likely facilitate the development of β-cells manufacture techniques. The aim of the present study was to investigate the pancreatic and duodenal homeobox 1 (pdx-1) gene expression into DTZ-stained cellular clusters originating from emberyonal carcinoma cells. This implemental -fundamental study was accomplished on differentiation of stem cells into insulin producing cells (IPCs). The conditioned medium of cultured pancreas from one-week newborn mouse was used for differentiation of P19. EBs formed by a 24h suspension culture of P19 EC cells. In order to induce the difrentiation،، different concentrations of conditioned medium (25%، 50%، 75% and 100%) were added to culture medium. Dithizone staininig was used for detecting the differentiated cells derived from EBs in vitro. Insulin-proinsulin production and insulin receptor beta were determined by immunofluorescence. The expression of pdx-1 gene was also analyzed by reverse transcriptase-polymerase chain reaction. Data were analyzed using one way ANOVA and Dunkana test. Differentiated cell clusters appeared after approximately 7 days induction. The peak response of differentiation was at the concentration of 50% conditioned medium. Expression of pdx-1 gene was observed in the differentiated cell clusters. Expression of insulin-proinsulin and insulin receptor beta markers in the differentiated cells was confirmed by immunofluorescence. The pancreas conditioned medium could influence P19 cells differentiation into insulin producing cells. So findings of this study could facilitate producing the β-cells.