Evaluation of a Multiwell Perfused Bioreactor System for the Long-term Culture of Mouse Hepatocytes

Liver is the primary organ for xenobiotic metabolism and in vitro models that faithfully recapitulate its function are needed for medium/high-throughput toxicity studies. We evaluated a three-dimensional multiwell perfused bioreactor system as a model of murine liver. System parameters (flow rate, seeding procedure and cell density, scaffold matrix, and media composition) were varied to identify optimal conditions for mouse hepatocytes. Biochemical assays (pyruvate, lactate, urea, albumin, and intracellular ATP) indicated that the three-dimensional system improves functionality of hepatocytes cultured for 3--5 days. We also tested hepatocyte's basal and inducible metabolism using model toxicants acetaminophen and WY-14,643. Finally, we cultured non-parenchymal cells with mouse hepatocytes to determine whether it may improve the functionality of cells. This work demonstrates that the three-dimensional multiwell perfused bioreactor seeded with mouse hepatocytes is a useful model for short-term studies; however, improvements in the system are needed for it to become a promising sub-chronic model.