Figure 2.

TGFB Signals from the Mesenchyme and Endothelium Are Candidate Regulators of O₂-Dependent Hepatocyte Differentiation in Liver Buds

(A) Phase-contrast and confocal images of hiPSC-LBs cultured for 1 (phase) or 5 (confocal) days (green: eGFP-iPSC-DE cells [AAVS1:EGFP]; red: KO1-HUVECs [MSCV-KO1]; no label: MSCs; scale bar, 250 μm).

(B) Boxplots of TGFB family gene expression in hiPSC-LBs cultured for 5 and 15 days. The error bars represent the maximum and minimum values; n = 9 (day 5) and 10 (day 15) independent experiments; ^∗p < 0.05 and ^∗∗p < 0.01 versus Excess-hypoxia; ^§p < 0.05 versus Ambient.

(C) Left: tSNE of single-cell transcriptomes from human fetal (two donors, gestation weeks 10.5 and 17.5, 238 cells) and adult (three donors, age 21–65, 256 cells) liver samples (modified from Camp et al., 2017). Right: violin plots of single-cell transcriptomes, showing the distribution in the human liver.

(D) Gene expression of each cell lineage in hiPSC-LB. hiPSC-LBs were cultured on Ambient group for 2 days. Then, dissociated eGFP-DE cells, KO-HUVECs, and unlabeled-MSCs from hiPSC-LBs were separated by FACS analysis using fluorescence labels. Gene expression of these cells was analyzed (mean ± SD; n = 8 independent experiments; ^∗p < 0.05 versus hiPSC-DE cells).