Optimising liver equivalents to model liver fibrosis

Zyoxel optimised the LiverChip with a Massachusetts Institute of Technology research group and Pfizer. The LiverChip is designed to mimic the hepatic architecture and is perfused and oxygenated by microfluidic flow under conditions akin to normal liver physiology. Cultures are contained within individual chambers, providing the "controllability" to accurately manipulate culture conditions to address the experimental question. Hepatocytes are viable and retain phase I and II metabolising enzyme expression and functionality for over 2 weeks. Published work revealed that the system can be used to co-culture hepatocytes and liver sinusoidal endothelial cells. In this pilot grant we will receive training from Zyoxel to establish the system in Newcastle, maximising our chance of success. The first objective is to optimise the seeding protocol to recreate a fibrotic liver, isolate hepatocytes and hepatic stellate cells (HSC) from rat liver, co-culture hepatocytes with increasing numbers of HSC, and assess culture viability and ask if co-culture causes HSC to HM activation or fibrosis using histological and biochemical readouts. The second objective is to co-culture hepatocytes with activated HM and monitor fibrogenesis. Once optimised in rats, the protocol will be translated for use with primary human cultures. The final objective is to create normal and fibrotic liver chips to be treated with or without proven anti-fibrotics (angiotensin inhibitors or 5HT2BR antagonist) to validate the system. Anti-fibrotic effects and compound toxicity will be assessed.