Production of albumin and urea by mouse hepatocytes on polyurethane foaming (PU) membranes coated with and in the absence of extracellular matrix (ECM) proteins was investigated in serum-free media for 7 days. The, amount of ECM proteins (collagen and fibronectin) coated on PU membranes was observed to decrease with an increase of time when ECM-coated PU membranes were immersed in culture media for 7 days. Hepatocytes cultured on PU membranes in serum-free media showed higher albumin and urea production than those cultured on polystyrene tissue culture flasks during a 7-day incubation. Three-dimensional culture of hepatocytes in ECM-coated PU foaming membranes was found to be more effective in the production of albumin and maintenance of hepatocyte viability than two-dimensional culture of hepatocytes. The viability of hepatocytes on laminin, vitoronectin and collagen coated PU membranes was found to be higher than that on a polystyrene tissue culture flask and non-coated PU membranes after a 7-day incubation. The effect of ECM proteins in ECM-coated PU membranes on hepatocyte function (albumin and urea production, as well as cell viability) was observed in hepatocytes cultured in serum-free media. (c) 2006 Elsevier B.V. All rights reserved.