Characterization of a perfused 3D liver bioreactor

Abstract

The liver is the most important site of drug and nutrient metabolism in the body, and we desire an accurate in vitro model that allows us to perform long term drug and metabolism studies. To this end of developing an assaying tool, I used an existing multi-well bioreactor that allows for formation of perfused, three dimensional tissue structures, and began characterization of tissue behavior over time. One issue in the multi-well bioreactor is the unknown profile of cell retention over time, which is an important specification for normalizing data from drug metabolism studies. Number of cells can be indirectly assessed by measuring total protein or RNA levels when direct counting is problematic. To the end of comparing these methods, an additional goal of this thesis was to develop a protocol to measure both protein and RNA levels from a single sample using the commercially available reagent RNAlater. RNAlater was shown, however, to be incompatible with certain existing protocols for isolating both protein and RNA.