Table 1.

Cellular models to study NAFLD in vitro.

Cellular System	NAFLD Induction	NAFLD Outcome	Observations	Ref.
2D Monocultures
PHH	NEFA	Steatosis, ER stress	Lipid accumulation, apoptosis, activation of autophagy (IRE1a), and lipid metabolism (ATF6a)	[41,42]
HuH7	NEFA	Steatosis, oxidative stress, inflammation	Lipid accumulation, apoptosis, expression IL-6, IL-8, TNFα, increased ROS, TGFB-1, TGFB -2, VEGF2	[44]
HepG2	NEFA	Steatosis, oxidative stress	Lipid accumulation, increased ROS, mitochondria changes (ATP levels, mitofusin-2 expression), impaired cholesterol efflux, and ABCA1 expression	[45,46]
	Endocrine disruptors	Steatosis, oxidative stress, lipoperoxidation, blocking autophagy	Lipid accumulation, TBARS expression, accumulation of autophagosomes, decreased SQSTM1/p62 degradation	[48,49]
	Drugs	Steatosis, oxidative stress, blocking autophagy	Lipid accumulation, increased lipogenesis (SREBP1c) and triglyceride formation (DGAT1), ROS generation, decreased SQSTM1/p62 degradation	[50,51,52]
L02	NEFA	Steatosis	Lipid accumulation, up-regulation of relevant cholesterol synthesis genes	[80]
	Valproic acid	Steatosis, oxidative stress	Lipid accumulation, decreased GSH level, increased MDA and ROS levels	[53]
HepaRG	Drugs	Steatosis	Decreased β oxidation, expression of enzymes involved in lipogenesis or decreased proteins involved in VLDL secretion	[56]
Upcytes	Drugs	Steatosis, oxidative stress	Lipid accumulation, decreased FOXA1 expression, increased ROS	[81]
2D cocultures
HuH7 and LX2	NEFA	Steatosis, HSCs activation	Lipid accumulation, α-SMA expression	[58]
AML12 and HSC	NEFA	Steatosis, oxidative stress, HSCs activation	Lipid accumulation, ROS induction, decreased CAT, SOD, and GPx, expression of profibrotic molecules (α-SMA, Col I, MMP-2, MMP-9, fibronectin)	[59]
PHH and KC	NEFA	Steatosis, inflammation	Lipid accumulation, expression of lipogenesis enzymes (FASN, SREBP1c),expression of TNFα, IL-1β, IL6	[61]
3D models
PHH	NEFA and insulin	Steatosis, insulin resistance	Lipid accumulation, increased expression of PCK1 and PDK4, and reduced GSK3β phosphorylation	[65]
HepG2 and LX2	NEFA	Steatosis, fibrosis	Lipid accumulation, Col1A1 expression	[66]
3D InSightTM	NEFA	Fibrosis, inflammation	Expression of collagen genes, fibronectin, α-SMA, IL-8 expression	[67]
PHH, HSC and macrophages	NEFA, insulin and glucose	Steatosis, insulin resistance, inflammation, fibrosis	Lipid accumulation, increased TAG, DAG and CE and PCK1 expression, reduced Akt phosphorylation, expression of IL-8, IL-6, and CXCL10, expression of TGF-β, OPN and α-SMA	[70]
PHH, HSC, LEC and KC	NEFA, TNFα and glucose	Steatosis, inflammation, fibrosis	Lipid accumulation, expression of IL-6, CXCL8, CXCL10, expression of MMP2 and MMP9	[71]
Liver-on-a-chip
HepG2	NEFA	Steatosis	Lipid accumulation, increased TAG	[75]
PHH, HSC, KC, and LSEC	NEFA and LPS	Steatosis, liver injury, fibrosis, inflammation	Lipid accumulation, ballooned hepatocytes, increased Caspase 3, expression of α-sma, col1a, timp-1, tgf-β and opn, increases in tnf-α, mip1a, and mcp1	[77]
HepG2 and gut cells	NEFA	Steatosis	Lipid accumulation	[78]