Table 4.

NAFLD, ALD, and DILI studies with organoid models

Reference	Liver disease	Culture approach	Study highlight
Ouchi et al. [58], 2019	NAFLD	iPSC-derived multi-cellular organoid	PSC-derived hepatic organoids containing both parenchymal and non-parenchymal cells types.
			When treated with free fatty acids, organoids recapitulate the main features of NAFLD, including steatosis, inflammation, and fibrosis.
Collin de l’Hortet et al. [180], 2019	NAFLD	iPSC-derived hepatocytes co-culture with mesenchymal cells, fibroblasts, and macrophages in decellularized rat liver	One of the largest organoids generated, containing multiple liver cell types. Tissue-like organoids enabled NAS scoring similar to patient biopsy samples.
			A peristaltic pump delivers nutrients and NAFLD-inducing agents to the core of the large organoid to achieve the manifestation of phenotypes throughout the tissue-like culture.
Kimura et al. [181], 2022	NAFLD	iPSC-derived multi-cellular organoid	En masse generation of liver organoids from 24 iPSC cell lines in a single matrigel droplet enables high throughput parallel screening for NAFLD phenotype.
			The high throughput genotype-to-phenotype correlation screen enables rapid validation and potential discovery of functional SNPs.
Ramli et al. [59], 2020	NAFLD	iPSC-derived hepatobiliary organoid culture	Hepatobiliary organoids comprise a bile canaliculi network extending from the hepatocyte core to the peripheral cholangiocyte cysts.
	DILI		The bile canaliculi system can model the cholestasis-inducing side effects of troglitazone.
			The hepatobiliary organoids treated with free fatty acids developed NAFLD phenotypes, including ductular reaction and diminishing bile canaliculi network.
McCarron et al. [140], 2021	NAFLD	ICO culture	NASH patient-derived ICO organoids exhibit increased expression of inflammation-associated genes and other genes associated with fibrogenesis and tumorigenesis.
			Idiopathic responses were observed across organoids generated from different patients.
Wang et al. [182], 2019	ALD	PSC-derived bipotent epithelial organoid	PSC-derived expandable hepatic organoids (EHO) can differentiate into functional hepatocytes or bile duct cells.
			Co-culture and differentiation of EHO with mesenchymal cells generate functional liver organoids that can model ALD pathophysiology after ethanol treatment, including oxidative stress, steatosis, inflammation, and fibrosis.
Shinozawa et al. [57], 2021	DILI	iPSC-derived hepatocyte organoid	Organoids comprise mainly hepatocytes with bile canaliculi-like architecture that supports unidirectional bile acid transport.
			The authors developed an oOrganoid-based drug toxicity assay with multiple readout measurements for cell viability, cholestasis, and mitochondrial toxicity. The organoid model showed high predictive value for 238 clinically available drugs.
			The study further demonstrated the organoid models’ utility for evaluating the pharmacogenomics effect of Bosentan-induced Cholestasis.

NAFLD, nonalcoholic fatty liver disease; ALD, alcoholic liver disease; DILI, drug-induced liver injury; PSC, pluripotent stem cell; iPSC, induced pluripotent stem cell; NAS, NAFLD activity score; SNPs, single-nucleotide polymorphisms; GCKR, glucokinase regulator; NASH, nonalcoholic steatohepatitis.