A high density culture of hepatocytes using a reticulated polyvinyl formol resin.

To enable high density culture of hepatocytes to be used as a hybrid artificial liver support or bioreactor system, collagen coated reticulated polyvinyl formal (PVF) resin (a filter material with a porosity of more than 80%) was used for the primary culture of hepatocytes. Stationary and perfusion culture experiments using PVF resin and monolayer culture were performed as control experiments. Due to the porous structure of the substrate material, hepatocytes were able to penetrate the reticulated pores of the PVF resin, and adhere to and spread on its surface. The densities of hepatocytes attained with PVF were about 10 times as high as those in the monolayer culture using conventional collagen coated Petri dishes. Hepatocytes immobilized in the PVF resin showed viability, as assessed by glutamic pyruvic transaminase (GPT) activity in the medium. Perfusion culture with PVF resin showed stable, high level metabolism, ammonium removal, and urea secretion comparable to the monolayer culture. It is concluded that perfusion culture with PVF resin is useful in attaining a high density culture of hepatocytes.