Fig. 3

Generation of the DhLOs model for drug metabolism. a Schematic of LOs generation from hiPSCs. b Representative morphology during DhLOs generation from normal and patient-derived iPSCs. Yellow arrowheads mark mesenchymal-like cells in patient-derived DhLOs, with an enlarged view provided in the right panel (yellow box). c mRNA expression of hepatocyte markers (ALB, AFP, CYP3A4) and cholangiocyte marker (KRT7) in normal and patient-derived DhLOs, compared to iPSCs. Data are mean ± SEM (n > 4 per group). d Immunofluorescence images of hepatocyte marker ALB and cholangiocyte marker KRT7 in DhLOs models. White arrowheads indicate cystic structures in patient-DhLOs. e Quantification of secreted ALB, ALT, and AST levels in EhLO and DhLOs. Data are mean ± SEM (n > 3 per group) (f) Enzyme activity of the CYP family in DhLOs with and without inducers (100 μM Omeprazole for CYP1A2 induction, 20 μM Rifampicin for other enzymes). RLU, relative luminescence unit. Data are mean ± SEM (n = 4 per group) g Fluorescence images of bile canaliculi-like structures stained with CLF in DhLOs, with an enlarged view of the left panel (white box) h mRNA expression of hepatic influx (top) and efflux (bottom) transporters in EhLO and DhLOs, compared to iPSCs. Schematic of hepatic drug transporters (lower left), created using Mind the Graph. Data are mean ± SEM (n > 4 per group). i Immunofluorescence images of influx transporter NTCP and efflux transporters (MRP2, BSEP, MRP4) in DhLOs. Data are mean ± SEM (n = 6 per group). Statistical comparisons were made using two-way ANOVA with Tukey’s test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001