Culture of primary rat hepatocytes within porous chitosan scaffolds

Chitosan is considered to be a very promising biopolymer for various biomedical and pharmaceutical uses because of its nontoxic and biocompatible natures (Chandy T, Shama P. Biomater Artif Cells Artif Org 1990;18:1-24). In this study, we prepared porous chitosan scaffolds by lyophilization of chitosan solution. The scaffolds were modified with water-soluble polyanionic species such as alginate and heparin. The pore structures of these scaffolds were viewed via light and scanning electron microscopy. The scaffolds prepared have a high porosity of approximately 90% with mean pore sizes from 50 to 200 microm. They were used as substrates for hepatocytes culture. The cell attachment ratio was much higher than on monolayer membrane and hepatocytes exhibited a round cellular morphology with many microvilli evident on the surface of the cells. Metabolic activities of the cells were evaluated in terms of albumin secretion and urea synthesis. It was found that hepatocytes cultured on the modified scaffolds showed an increase in albumin secretion during the first 4 days and were more stable than those on monolayer membrane and nonmodified scaffolds. Therefore, primary rat hepatocytes cultured on modified scaffolds would be beneficial to liver assist device.