Fig. 5

Robust induction of the hepatocyte program in iHDF-T. iHDF-T maintain inducibility after more than 80 population doubling without losing hepatic phenotype in diHLC-T. A Volcano plot of all genes expressed in PD^low and PD^high diHLC-T (three replicates each) by RNAseq. Genes with higher expression in PD^high diHLC-T are depicted as LOG₂(FC) > 0. Plotted genes were expressed in all samples of at least one group (absolute reads > 10). Red, green and yellow dots correspond to genes differentially expressed defined as FDR < 0.05 and fold change > 2. Green dots represent genes encoding hepatocyte-secreted plasma proteins. Yellow dots represent genes encoding hepatocyte-relevant secreted glycoproteins such as decorin (DCN) or fibromodulin (FMOD). A complete list of differentially expressed genes is included in Additional file 2: Table S4. B GO molecular function enrichment within differentially expressed genes. Dashed red line marks FDR = 0.05. C Comparison of relevant CYP450 activities between PD^low and PD^high diHLC-T. Activities were quantified as in Fig. 4E. D Induction of CYP450 activities by phenobarbital and rifampicin. Activities were quantified as in Fig. 4E. E Volcano plot of metabolomics features (m/z-Rt) quantified in PD^low and PD^high diHLC-T (six replicates each) by liquid chromatography-TOF. Features with LOG₂(FC) > 0 are more abundant in PD^high diHLC-T. Plotted features were expressed in all samples of at least one group. Red dots correspond to features differentially abundant defined as FDR < 0.05 and fold change > 2. F HCS analysis of amiodarone incubation at 1–10-100–1000 µM in PD^low and PD^high diHLC-T (six replicates each). *p < 0.05. G Relative mRNA level of multiple hepatic genes in P5E10 clone (PD30-40). Values correspond to the average plus standard deviation of two different experiments (four samples each) determined by RT-qPCR and expressed relative to PD^low (P3G4). For comparison, relative mRNA levels of PD^low (PD30-40), PD.^high (PD100-110) and PD125 of P3G4 clone are depicted in Additional file 1: Fig. S4