Figure 4.

Evaluating the role of growth factor, matrix, and amino acid in the induction of early recovery in hepatocytes cultured in 3-D configuration. A) Hepatocyte albumin secretion on d 4 of culture following addition of neutralizing antibodies against HGF, IL-6, or TNF-α to coculture of hepatocytes and endothelial cells (black bars) or exposure of monocultures of hepatocytes to culture medium supplemented with recombinant HGF (200 ng/ml), IL-6 (1 ng/ml), or TNF-α (0.5 ng/ml) (white bars). B) Albumin secretion by hepatocytes on d 4 of culture following stimulation with 75% endothelial cell-conditioned medium compared to conditioned medium treated with MMP-3 (1 μg/ml); albumin secretion by hepatocytes on d 4 of culture in hepatocyte culture medium (0%) supplemented with laminin (10 μg/ml), fibronectin (10 μg/ml), or MMP-3 (1 μg/ml). Results are normalized to hepatocytes cultured in conditioned medium (75%).**P < 0.01 vs. conditioned medium; ^##P < 0.01 vs. control medium. C) Dose-dependent inhibition of albumin secretion by hepatocytes (d 4) exposed to 75% endothelial cell-conditioned medium supplemented with cis-hydroxyproline (HP). Results are normalized to hepatocytes cultured in conditioned medium (75%). Addition of exogenous proline dramatically increases the albumin secretion of nonstimulated hepatocytes. **P < 0.01 vs. conditioned medium; ^##P < 0.01 vs. control medium. D) Dose-dependent inhibition of albumin secretion (d 4) in cocultures of hepatocytes and endothelial cells stimulated with control medium (0%) supplemented with cis-hydroxyproline (HP). Results are normalized to cocultures exposed to control medium (0%). **P < 0.01 vs. control medium.