A high-density culture of hepatocytes using porous substrate for use as a bioartificial liver

Abstract

High-density, large-scale culture of hepatocytes is a key requirement in the development of a bioartificial liver that can replace liver functions in patients with severe liver insufficiency. We have applied a porous polymer, polyvinyl formal (PVF) resin, as a cell-supporting material for hepatocyte culture. We evaluated the performance of the culture system using PVF resin under three different culture conditions: a shake culture on conventional dishes, perfusion culture with sheet-shaped PVF, and a packed-bed-type module. Among them, the packed-bed reactor using PVF resin enabled high-density culture of hepatocytes (2×107 cells/cm3-PVF). The hepatocytes immobilized in the PVF resin maintained satisfactory metabolic functions (ammonium metabolism and albumin secretion) comparable to those of the monolayer dish cultures. Furthermore, by maintaining dissolved oxygen concentration at a relatively high level (260–460μM), the metabolic functions of the hepatocytes were improved. It was concluded that the packed-bed reactor using PVF resin is a promising system for developing a bioartificial liver using hepatocytes.