Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens

Generation of human hepatocytes-derived liver progenitor-like cells in vitro. a Overview of the protocol used to convert PHCs into HepLPCs. b Light microscopy images of PHCs cultured in HGM or TEM at day 0 and day 10. Scale bars, 200？μm. c QPCR analyses for the expression of the indicated genes during TEM culture from day 0 to day 10. FLCs, fetal liver cells, 5 gestational weeks (one-way ANOVA with Dunnett correction for multiple comparisons, n？=？4 donors, *P？<？0.05, **P？<？0.01, ***P？<？0.001). d Immunofluorescence analyses demonstrating the expression of ALB and CK19. Scale bars, 50？μm. e Flow cytometric analysis showing the proportion of ALB or CK19 positive-cells in the populations. Blue, positive-cells; red, negative controls. f Growth curves illustrate the number of cells counted per well at different time points. Results expressed as mean？±？s.d. of three independent cultures derived from three donors. g Doubling time calculated for three donors. Error bars represent s.d.; n？=？3. h Edu incorporation is detected at passage 10. Scale bar, 50？μm. i Representative karyotype images of three independently established HepLPCs. Two lines maintained normal diploid karyotypes, whereas the third was partly triploid for Chromosome