. 2021 Dec 30;11(1):86. doi: 10.3390/antiox11010086

Table 2.

Pluripotent stem cell-derived cell models to study NAFLD.

Cell Model	HLCs Characterization	NAFLD Induction and Model Observations	Ref.
ESC line H1 and iPSCs from healthy donors	Polygonal shape, alb⁺, ecad⁺, hnf-4α⁺, urea synthesis, CYP3A4/3A5/3A7 activities, release of indocyanine dye	NEFA for 48 h. Lipid accumulation. Up-regulation of lipid metabolism regulators PPARα and PLIN2. Down-regulation of certain microRNAs (microRNA hsa-miR-122 and hsa-miR-106b).	[19]
HLCs from NAFLD donors (distinct grades of steatosis)	Polygonal shape, alb⁺, afp⁺, ecad⁺, hnf-4α⁺, a1at⁺, and ttr; low CYP3A4 expression	NEFA for several days. Lipid accumulation with a donor-specific pattern. Increased PLIN2 expression with differences between donors. Low expression of the genes associated with FGF21 signaling, lipid and cholesterol biosynthesis, and gluconeogenesis, with a low expression of CPT1A in high steatosis lines. AdipoRon effect on metabolism, transport, and signaling pathways.	[103]
iPSCs-Hep from female healthy donors	Polygonal shape, alb⁺, afp⁺, pou5f1⁺, hnf-4α⁺, expression of phase I, II, and III enzymes	NEFA and TAG for 18 h. PA dose-dependent lipid accumulation. Lipid accumulation exacerbated by TAG treatment, which induces ER stress by UPR dysregulation. Up-regulation of the genes related in fat storage in lipid droplets. Down-regulation of β-oxidation genes (ACADM) with FA-TAG treatment. Reduced TAG accumulation after inhibition of ER stress by therapeutic molecules tauroursodeoxycholic acid or obeticholic acid.	[94]
iPSC lines FSPS13B and A1ATDR/R obtained by CRISPR/Cas9 technology	Polygonal shape in HLCs alb⁺, a1at⁺, hnf-4α⁺ Detected CYP3A4 activity	NEFA for 7 days. HLCs-PNPLA3^KO and HLC-PNPLA3^I148M do not activate UPR markers (BIP, GADD34, CHOP, and PERK) with PA treatment, which indicates lipid-associated ER stress alterations. Lower levels of several β-oxidation gene expressions. Down-regulation of the genes implicated in drug detoxification, glucose metabolism and cell stress, but sensitivity to insulin remaining. HLCs-PNPLA3^I148M recapitulate the main PNPLA3-associated NAFLD features.	[96]
Female H9 ESCs	Polygonal shape, alb⁺, hnf-4α⁺ Detected secreted albumin and CYP1A2 and CYP3A4 activities	Lactate, pyruvate and octanoate for 48 h or 96 h. Increased lipid accumulation. Tricarboxylic acid cycle dysregulation and altered expression of related enzymes. Alteration of β-oxidation and oxidative phosphorylation. Up-regulation of lipid vesicle transport proteins PLIN1, PLIN2 or APOA4, and gluconeogenesis genes. Transcriptional dysregulation in insulin resistance mediators. Induction of oxidative stress.	[97]
iPSCs from NASH and healthy patients	Polygonal shape with bile canaliculi formation. Hepatic markers (aat⁺, alb⁺), mRNA expressions of CYP3A4, CYP3A7, SERPINA, ASGR1, and ALB	NEFA for 24 h. Increased dose-dependent intracellular lipid accumulation. NAFLD patient derived HLCs organoids spontaneously accumulate lipids. HLCs were able to successfully integrate into 3D liver organoids with macrophages, MSCs and endothelial cells allowed NAFLD to be more accurately modeled.	[93]
3D coculture (HLCs and other cells in decellularized rat liver)	Polygonal shape, alb⁺, hnf-4α⁺ mRNA expressions of the enzymes, transporters, hepatic nuclear receptors, and transcription factors involved in liver metabolism	Linoleic acid and OA in the presence of DOX for 2–4 days. Increased lipid accumulation, lipid peroxidation levels, and total cholesterol levels. Increased mRNA expression of the genes implicated in de novo lipogenesis (SREBP1c). Decreased mRNA expression of β-oxidation key modulators (PPARα and PGC1α). Bioengineered tissue exhibits steatosis and expresses pro-inflammatory markers.	[92]
3D organoids (HLCs and CLCs deriving from PSCs)	alb⁺ (HLCs), ck7⁺ (CLCs), mRNA expression of proteins related to bile acid synthesis/secretion, cholesterol, fat and carbohydrate metabolism, drug detoxification, and hepatic-specific transcription factors	NEFA exposure. Increased lipid accumulation, ROS levels, and lipid peroxidation. Increased expression of the genes related to lipid and carbohydrate metabolism. Bile canaliculi network disruption.	[95]