Modeling pathogenesis and progression of metabolic dysfunction-associated steatotic liver disease and therapeutic drug screening using hESC-derived mature polarized hepatocyte organoids

• Established a novel in vitro MASLD model using hESC-derived mature polarized hepatocyte organoids (P-hep-orgs) treated with free fatty acids, recapitulating key pathological hallmarks including disrupted metabolism, oxidative stress, and loss of polarization. • Transcriptomic analysis revealed significant molecular overlap (581 differentially expressed genes) between FFA-treated P-hep-orgs and human MASH liver tissues, supporting the model's clinical relevance. • Demonstrated the utility of the P-hep-org MASLD model for therapeutic drug screening by evaluating known antioxidant and lipid-lowering agents, such as Vitamin E, which alleviated lipid accumulation and oxidative stress. • The P-hep-org model overcomes limitations of existing animal and 2D culture models by providing a physiologically relevant, polarized, and functional human hepatocyte platform for studying MASLD pathogenesis and drug discovery.