Influence of pancreatic islets on growth and differentiation of hepatocytes in co-culture.

Improvement of cell culture conditions in hepatic tissue engineering may permit cell/tissue banking and the generation of liver tissue equivalents for transplantation. In these systems, continuous hepatotrophic stimulation is still necessary. We investigated the stimulatory effects of pancreatic islets on hepatocytes in co-culture and characterized the stimulatory mechanisms. Hepatocytes and pancreatic islets were harvested from Lewis rats. Cells were cultured on collagen dishes either with nonstimulated media (controls and co-cultures with low or high islet rate) or stimulated media (controls and co-cultures). To characterize stimulatory mechanisms, additional co-cultures with membrane separation, with antiinsulin, antiglucagon, and with both antibodies were examined. Hepatocyte numbers, albumin secretion rate by enzyme-linked immunoadsorbent assay, and monoethylglycinxylidid biotransformation values by fluorescence polarization immunoassay were assessed. A radioimmunoassay measured insulin and glucagon concentrations. In groups with nonstimulated media, cell number was higher in co-cultures with low islet rate, and albumin secretion rate was increased in co-cultures with high islet rate compared to controls. MEGX biotransformation was decreased in co-cultures. In groups with stimulated media, co-culture had no impact on cell number or albumin secretion rate. Hepatocyte numbers and albumin secretion rates were not changed in co-cultures after membrane separation. Islet effects on hepatocytes were reduced in co-cultures with antiinsulin, antiglucagon, or both antibodies. Pancreatic islets provide stimulation for hepatocytes in vitro. Islet effects were mediated by soluble factors, and are dependent on insulin and glucagon. These results permit further investigations towards three-dimensional transplantable hepatocyte-islet devices for continuous in vitro and in vivo stimulation.