Human Di-lineage Hepatic Spheroids Reproduce Key Features of MASLD : Cell Mol Gastroenterol Hepatol | May 2026

Introduction

Metabolic dysfunction-associated steatotic liver disease affects nearly one-third of the global population and spans a disease spectrum from steatosis to steatohepatitis, fibrosis and cirrhosis. Despite major advances in clinical hepatology, the molecular mechanisms driving transition from simple steatosis to progressive fibrotic liver injury remain incompletely understood.

Problem Statement

Existing in vitro liver disease models—including immortalized hepatocyte systems and induced pluripotent stem cell organoids—often fail to fully reproduce the complex metabolic and fibrotic interactions observed in human MASLD. More physiologically relevant human models are needed to better study disease biology and evaluate antifibrotic therapies.

Summary

This translational study developed a human di-lineage hepatic spheroid platform combining primary human hepatocytes and hepatic stellate cells in physiologic proportions to model steatotic liver disease and fibrosis. Exposure to free fatty acids and transforming growth factor-β1 successfully induced hallmark features of MASLD and MASH, including intracellular lipid accumulation, increased collagen deposition, impaired ApoB100 secretion and development of a profibrotic secretory profile characterized by reduced matrix metalloproteinases and increased tissue inhibitors of metalloproteinases. Multiomic integration using transcriptomics and proteomics demonstrated strong concordance between gene and protein expression and identified activation of extracellular matrix remodeling and TGFB signaling alongside suppression of metabolic and cholesterol pathways. Importantly, the molecular signatures observed in the spheroids closely mirrored fibrosis-associated gene expression patterns in human liver tissue from the GepLiver database, supporting translational relevance of the model. Key fibrosis-associated markers such as COL1A1, ACTA2, TGFBI and PLOD2 were strongly upregulated, whereas lipid metabolic regulators including APOA2 and FABP1 were suppressed, reflecting simultaneous fibrotic remodeling and metabolic dysfunction. The spheroids also demonstrated pharmacologic responsiveness to resmetirom and obeticholic acid, both of which reduced lipid accumulation and collagen expression, paralleling observations from clinical studies. Although limited by exclusion of additional liver-resident immune and endothelial cell populations, this simplified human primary-cell platform provides a robust and physiologically relevant experimental system for studying MASLD progression, identifying therapeutic targets and evaluating antifibrotic interventions.