Figure 3.
Differentiation of Hepatocytes from hiPSC-Derived CPM+ LPCs
(A) Schematic image of hepatocyte-like cell differentiation from CPM+ LPCs.
(B–D) The morphology of CPM+ LPCs (upper) and CPM+ hepatocyte-like cells (lower) was investigated by microcopy. (B) Phase contrast images. CPM+ LPCs exhibit light cytoplasm and indistinct cell borders. CPM+ hepatocyte-like cells exhibit cobblestone-like morphology with binucleation. (C) Immunohistochemistry for AFP (red), ALB (green) and HNF4α (gray). Nuclei were counterstained with Hoechst 33342 (blue). (D) PAS staining showed accumulation of glycogen. Scale bars, 100 μm.
(E) Uptake of DilAcLDL (red) in CPM+ hepatocyte-like cells. Scale bar, 100 μm.
(F) qRT-PCR analysis of various CYP450s mRNA levels. The results are shown as the mean ± SEM of six independent experiments. (each experiment contains two technical replicates). iPSC-Heps was used as a control. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
(G) Secretion of ALB and urea. The results are shown as the mean ± SEM of seven independent experiments. iPSC-Heps was used as a control. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
(H) Relative CYP3A4 activity. The results are shown as the mean ± SEM of at least three independent experiments. Treatment with 10 μM of rifampicin (Rif) for 72 hr. iPSC-Heps was used as a control. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.