Figure 5: Development of fetal liver parenchymal and non-parenchymal cells.

(a) CEBPα⁺ hepatocyte-like cells and CD31⁺ CD34⁺ vascular networks on day 14. (b) DES⁺ stellate-like cells (c) CK19⁺ AAT⁺ hepatocyte-like cells with CD146⁺ vascular-like structures. Scale bar, 200 μm. (d) Heatmap shows upregulation of hepatic genes between days 5 and 10. (UBC and EEF1A1 as used as control/housekeeping genes). (e) CK7⁺ bile duct-like channels develop within fetal hepatocyte-like cells; xz and yz slices on the left and bottom. Scale bar, 200 μm. (f) Further maturation of liver-like phenotype as evidenced by increasing albumin production (ELISA) and (g) albumin⁺ cells. Data are mean±s.e.m., n=3 per group. phAlb_mKate2 is a lentivirally integrated reporter where a short human albumin promoter drives expression of a red fluorescent protein (mKate2). Scale bar, 200 μm. (h) Confocal imaging and three-dimensional reconstruction of AAT⁺ hepatocyte-like cells (red) with an apical layer of CD34⁺ endothelial-like cells (green). Scale bar, 50 μm.