Transcriptomic analysis of hepatocyte-like cells generated by doxycycline inducible expression of HNF4A, HNF1A and FOXA3

We have generated an immortal fibroblast-derived cell line without malignant traits capable of being efficiently and easily reprogrammable into hepatocyte-like cells under standard culture conditions by combining hTERT expression and a polycistronic vector inducible with doxycycline. Overall design: Human fibroblasts were immortalized with hTERT and infected with a lentiviral vector containing a polycistronic 2A cassette expressing HNF4A, HNF1A and FOXA3 under the control of Tetracycline Response Elements (iHDF-T). After single cell cloning, we selected a specific clone named P3G4. This clone was expanded for more than 80 population doublings (PD). We reprogrammed by doxycycline addition, low and high PD iHDF-T into hepatocyte-like cells (diHLC-T). RNA from PD low and high iHDF-T as well as diHLC-T were analyzed by RNAseq in triplicates. RNA isolated from 24hr-cultured primary hepatocytes from three different donors were used as controls.