Fig. 2: hiHepPCs are directly induced from HUVECs and differentiate into hepatocytes.

a GSEA of CEL-seq2 data for hiHepPC monolayer cultures at passage 8 and HUVECs was performed using the set of top 100 genes specifically upregulated in hHepPCs derived from hESCs¹³ or adult human livers¹⁴. b qPCR analyses of hHepPC marker genes were performed on total RNA obtained from HUVECs, three different hiHepPCs at passage 8 in monolayer culture, and human hepatocytes. All data were normalized with the values for hiHepPC-1 (monolayer), and the fold differences are shown. Data represent the mean ± SD (n = 3 independent assays). c Co-immunofluorescence staining of ALB with AFP was conducted for hiHepPCs induced from HUVECs at the indicated passage numbers (P). P1 and P1’ designate days 4 and 7, respectively, after the initial passage of transduced HUVECs. The upper and lower right graphs show the percentages of AFP⁺ cells in ALB⁺ cells and ALB⁺ cells in AFP⁺ cells, respectively, which were observed in individual cultures of hiHepPCs during passaging. Data represent the mean ± SD (n = 3 independent experiments). d Co-immunofluorescence staining of ALB with E-CAD, AAT, ASGPR1, or CYP3A4 and of AFP with HNF4A or CYP3A4 and immunofluorescence staining of CD31 were conducted for mock-infected HUVECs at passage (P) 1 and hiHepPC monolayer cultures at P6. Periodic acid–Schiff (PAS) staining and oil red O staining were also conducted for mock-infected HUVECs and hiHepPC monolayer cultures to visualize glycogen stores and lipid synthesis, respectively, in hiHepPC-derived hepatocytes. Additionally, ICG uptake and subsequent release by hiHepPC-derived hepatocytes are shown. DNA was stained with DAPI. Scale bars, 50 µm. Source data are provided as a Source Data file.