Fig. 1

Generation of a novel multicellular organotypic liver model. a Schematic diagram of hepatic spheroid generation and macroscopic images of 3D spheroids. The indicated numbers of HepaRG cells, HUVECs, and MSCs were self-assembled on a Matrigel bed in a conical tube. b Distributions of HepaRG cells (ALB), HUVECs (CD31), and MSCs (Desmin) within cryosections of 3D hepatic spheroids on day 12. The nuclei were counter-stained with DAPI (blue). c mRNA expression levels of the indicated hepatocyte-specific markers were determined in differentiated HepaRG cells cultured alone or with HUVECs and MSCs in 2D or 3D triple co-culture using real-time PCR on day 12. β-Actin expression was used as an internal control; n = 3. d ALB production and e CYP3A4 activity were determined in 2D triple co-culture or 3D hepatic spheroids on day 12, normalized to the ATP concentration; n = 3. *p < 0.05, **p < 0.01, and ***p < 0.001