HNF4α transduction and 5'azacytidine treatment of naïve rat liver epithelial cells (rLEC)

Neonatal liver-derived rat epithelial cells (rLEC) from biliary origin are liver progenitor cells that acquire a hepatocyte-like phenotype upon sequential exposure to hepatogenic growth factors and cytokines. Undifferentiated rLEC express several liver-enriched transcription factors including the hepatocyte nuclear factors (HNF) 3β and HNF6, but not the hepatic master regulator HNF4α. In this study, we investigated the impact of the ectopic expression of HNF4α in rLEC on both mRNA and microRNA level by means of microarray technology. In addition, we investigated the influence of subsequent DNA methyl transferase (DNMT) inhibition on the phenotype of undifferentiated rLEC by exposure to 5' azacytidine (AZA). rLEC were transduced with a lentiviral vector containing HNF4α and puromycin selected. In addition, cells were exposed to 20 µM 5'azacytidine for 7 days