Fig 1: Functional analysis of cultured induced pluripotent stem cell-hepatocyte-like cells (iPSCs-HLCs) on the dish and iPSCs-HLCs seeded into the scaffold. (A) Protocol for differentiation induction to hiPSC-HLCs. (B) Bright field and immunostaining images of hiPSCs-HLCs at day 25 of differentiation induction. Left to right: bright field, SOX17, human albumin, and AFP. Scale bar = 200 µm. (C) Comparison of ALB secretion level between cultured hiPSC-HLCs and primary human hepatocyte (PHH) on dishes. (D) Bioreactor for recellularization. (E) Comparison of CYP3A4 activity for the cultured hiPSC-HLCs in the scaffold and on the dish (mean ± SD, n = 3). (F) Comparison of relative CYP-related mRNA expression in the cultured hiPSC-HLCs in the scaffold and on the dish (mean ± SD, n = 3) (G) H&E staining of the recellularized scaffold. (H) Immunostaining images of the recellularized scaffold. Left: human albumin. Right: CYP3A4. Scale bar = 100 µm. (I) Renal subepithelial implantation of the recellularized scaffold (J) Histological images of the scaffold observed after 14 d of the implantation. Scale bar = 50 µm. (K) Immunostaining images of the implanted scaffold. Left: human ALB and pig CD31. Right: CYP3A4. Scale bar = 100 µm.