Stimulation of in vitro rat hepatocyte proliferation by conditioned medium obtained from an immortalized macrophage cell line

The hepatomitogenic effect of conditioned medium (CDM), obtained from the N-11 mouse macrophage cell line was analysed in rat hepatocyte primary cultures. CDM concentrations from 0.01% to 100% were used and the stimulating action in terms of mitotic index (MI) was evaluated. A clear mitogenic effect was observed only with concentrations higher than 10% with peak effects around 60%. Further increase in CDM concentrations resulted in an MI decrease, and at 100% CDM the effect was totally abolished. Tests addressed to identify the presence of hepatocyte growth factor (HGF) yielded negative results. In order to identify the mitogenic factor(s) involved, we tested CDM obtained after lipopolysaccharide (LPS) stimulation of N-11 cells. Comparison of the results obtained with untreated or LPS stimulated CDMs suggested that macrophage activation does not affect the release of hepatomitogenic activity. To further characterize this macrophage-derived activity, we checked whether CDM could interact with the mitogenic effects of epidermal growth factor (EGF). CDM (10 or 50%) showed no stimulatory effect to hepatocytes cultured in the presence of a maximally stimulatory concentration of EGF. Conversely, both CDM concentrations were able to increase the MI of hepatocyte cultures treated with a suboptimal dose of EGF. These results suggest that macrophages release factor(s) which interact, in hepatocytes, with the EGF signal transduction mechanisms, or with the EGF receptor itself. Copyright (C) 1999 Elsevier Science Ltd.