Modelling fatty liver disease with mouse liver-derived multicellular spheroids

Area of Science:

• Hepatology and Toxicology
• In Vitro Disease Modeling

Background:

• Chronic liver disease, fibrosis, and cirrhosis pose significant global health challenges.
• Current in vitro models for liver disease lack reliability, hindering drug development.
• Primary cell-based multicellular liver spheroid (MCLS) cultures offer a promising alternative.

Purpose of the Study:

• To establish and characterize functional multicellular liver spheroid (MCLS) cultures using primary mouse cells.
• To assess the utility of MCLS cultures in modeling liver fibrosis and non-alcoholic fatty liver disease (NAFLD).
• To evaluate the efficacy of potential anti-NAFLD drugs in the established MCLS model.

Main Methods:

• Primary mouse hepatocytes, hepatic stellate cells, liver sinusoidal endothelial cells, and Kupffer cells were aggregated into spheroids using 96-well U-bottom plates.
• MCLS cultures were maintained for up to 17 days and characterized for cellular composition and function.
• Liver disease models were induced using acetaminophen (fibrosis) and free fatty acid/lipopolysaccharides (steatosis and fibrosis).
• The efficacy of Elafibranor, Lanifibranor, Pioglitazone, and Obeticholic acid in treating induced liver conditions was assessed.

Main Results:

• Functional MCLS cultures containing viable hepatocytes, quiescent stellate cells, fenestrated endothelium, and responsive Kupffer cells were successfully established.
• MCLS cultures recapitulated fibrotic responses to acetaminophen and displayed steatosis and fibrosis upon challenge with fatty acids and LPS.
• All tested drugs inhibited steatosis; Elafibranor and Lanifibranor demonstrated significant inhibition of fibrosis.

Conclusions:

• Primary mouse MCLS cultures serve as a robust in vitro model for acute and chronic liver diseases, including NAFLD.
• This model is suitable for evaluating the therapeutic potential of anti-NAFLD drugs, particularly their anti-fibrotic effects.
• MCLS cultures represent a significant advancement over traditional cell line mono-cultures for liver disease research.