Fig. 1

Functional characterization of human liver organoids derived from hPSCs. A Schematic representation of the differentiation method for human liver organoids. DE represents definitive endoderm. B Representative bright field image (4 × , 20 ×) of HLOs or 5F-HLOs at day 20. Scale bar, 100 μm, 50 μm, respectively. C, D The number and the albumin secretion level of HLOs or 5F-HLOs. E Expression level of representative genes in iPSC, HLOs and 5F-HLOs. Hepatic markers (ALB, CYP3A4, CYP2B6, CYP2E1), stellate cell markers (PDGFRβ, COL1α1), cholangiocyte markers (KRT19, SOX9), and kupffer cell markers (CD14, CD68). mRNA levels were assessed by qPCR and expressed as fold versus iPSC. n = 3 independent differentiations, *P < 0.05, #P < 0.05 versus HLO. F Immunostaining for Albumin (ALB), SOX9 and PDGFRβ in HLO and 5F-HLO. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. G CDCFDA uptake and release in HLO and 5F-HLO. Scale bar, 50 μm. H LDL uptake evaluated by DiI-LDL fluorescent staining in HLO and 5F-HLO (DiI-LDL in red, and DAPI in blue). Scale bar, 50 μm. I Glycogen accumulation evaluated by PAS staining in HLO and 5F-HLO. Scale bar, 50 μm. Values represent means with SEM. P values were assessed by unpaired, two-tailed Student’s t test (C, D), and one-way ANOVA with Dunnett’s multiple comparisons test (E)