Overcoming Mass Transfer Barriers in Sandwich Configuration for Primary Hepatocytes Culture

Abstract

This thesis explored two novel ways to encounter the inherent mass transfer barriers of conventional sandwich configuration for primary hepatocytes culture. Insufficient mass transfer in sandwich configuration is known to cause metabolite accumulation and limited nutrient access in the intra-sandwich environment. We successfully designed a novel bioreactor for perfusion sandwich culture with ability to effectively control the mass transfer efficacy. The bioreactor created two flow streams: a main flow stream flowing culture medium above the upper extra-cellular matrix support on porous membrane and a separate drainage stream created by channels directly connected to the intra-sandwich environment, facilitating the removal of the metabolites and supply of nutrients directly. Also we managed to improved the mass transfer efficacy by replacing the natural ECMs such as collagen, which is conventional used in forming sandwich configuration, with the synthetic polymers with controllable physical and chemical properties. An ideal synthetic sandwich configuration with better mass transfer efficacy and hepatocyte function was identified by overlaying a novel 3D monolayer developed on galactosylated PET film with RGD conjugated polyethylene terephthalate (RGD-PET) membranes.