Fig. 5: hiHepPC-derived cells functionally reconstitute hepatic tissues in vivo. | Nature Communications

Fig. 5: hiHepPC-derived cells functionally reconstitute hepatic tissues in vivo.

From: Direct reprogramming of human umbilical vein- and peripheral blood-derived endothelial cells into hepatic progenitor cells

Fig. 5

a Schematic diagram of the experimental procedure. hiHepPCs were marked by infection with a virus expressing enhanced green fluorescent protein (EGFP) before transplantation. PH partial hepatectomy. b Immunofluorescence staining of the thymidine analog 5-bromo-2’-deoxyuridine (BrdU) was conducted for the livers of control mice (No treat) and retrorsine-treated and untreated mice at day 2 after PH. The graph shows the percentages of BrdU+ cells observed in individual mouse livers. c Kaplan–Meier survival curves of the recipient retrorsine-treated hepatectomized mice after intrasplenic injection of human hepatocytes, cells dissociated from hiHepPC aggregates and hiHepPC monolayer cultures, HUVECs, or only PBS into the liver. Statistical analyses using the log-rank test revealed significant differences between the curves for HUVECs and hiHepPC aggregates or hepatocytes but not between those for hiHepPC aggregates and hepatocytes (P = 0.309). d Immunofluorescence staining of human AAT (hAAT) was conducted for recipient mouse livers 2 months after transplantation. The graph shows the percentages of hAAT+ cells observed in individual mouse livers. e Co-immunofluorescence staining of EGFP with hAAT, CYP3A4, E-CAD, human CK8/18 (hCK8/18), MRP2, or HNF4A and of hAAT with human ALB (hALB) were conducted for recipient mouse livers 2 months after transplantation of cells dissociated from hiHepPC aggregates. f, g The amounts of AST and ALT (f) and those of hALB (g) in the serum of recipient mice were measured 2 days and 2 months after transplantation, respectively. The sera of control mice (No treat) were measured as negative controls. h Schematic diagram of the experimental procedure. i Co-immunofluorescence staining of human CK19 (hCK19) with mouse CK19 (mCK19) or SOX9 was conducted for DDC-treated mouse livers 4 weeks after the last injection of cells dissociated from hiHepPC spheroids or human fetal cholangiocyte-derived spheroids. j Co-immunofluorescence staining of hALB with mouse Alb (mAlb) was conducted for retrorsine-treated hepatectomized mouse livers 2 months after transplantation of cells dissociated from hiHepPC aggregates or human hepatocytes. Statistical difference was determined by one-way analysis of variance followed by Tukey–Kramer test (b) or Dunnett’s test (d, f, g). Data represent the mean ± SD (n = 3 (b, d) or 4 (f, g) independent experiments). DNA was stained with DAPI. Scale bars, 50 µm. Source data are provided as a Source Data file.

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