Figure 3. Liver-specific gene expression was augmented in 3D HepaRG spheroids.

HepG2 and HepaRG cells were seeded into 96-well plates at 2000 cells/well (HepG2), 72000 cells/well (2D, HepaRG) or 24000 cells/well (3D, HepaRG) and subsequently cultured for up to 7 days. Cells were harvested at indicated days: Relative mRNA levels of genes involved in drug metabolism (A), gluconeogenesis (B), glycolysis (C), energetic lipid synthesis (D), bile acid metabolism (E) and lipoprotein metabolism (F) and levels of GAPDH (G) were determined by quantitative RT-PCR and were normalized to levels of 18s rRNA. All assays were performed in triplicate. Error bars indicate ± S.E.M. **P<0.01, compared with 2D cultures performed on each day.