Figure 2.

BALM Cell Repopulation

(A) Macroscopic appearance of a mouse liver scaffold during seeding and culture. Decellularized liver is flushed with media to identify well-perfused lobes, which are seeded on day 6; the white arrows indicate the visible pockets of DECs. Scaffold harvested at day 25 of differentiation loses transparency. Scale bar, 5 mm.

(B) DAPI nuclear staining (grey) show ECM fibres and absence of nuclei on a cryosection of an unseeded lobe (middle and left). hiPSCs-derived nuclei are present along scaffold fibres of a cryosection of a DECs seeded lobe at day 26 (right). DAPI intensity gains are identical between left and right image, middle image is intensity gain x4 of left image to visualize the ECM fibres. Scale bar 100 µm. (C) H&E staining (nuclei stained in purple and ECM in pink) also show ECM fibres and absence of cells on a cryosection of an unseeded lobe (top). hiPSCs-derived cells appear to be positioned along the scaffold fibres of a cryosection of a DECs seeded lobe at day 26 (bottom). Higher magnification images correspond to the delineated lower magnification images in the adjacent panel. Scale bar 100 µm.

(D) Ki-67 staining of a cryosection of a DECs seeded lobe at day 21 shows proliferating cells (top) and few apoptotic cells as shown by Cc3-positive staining (bottom). Nuclei stained with DAPI. Scale bar, 100 μm.

See also Figures S2 and S5 and Table S3.