Table 1 |.

Most commonly used in vitro and mouse and rat models of NASH

Platform	Description	Phenotype and relevance to human disease
In vitro
Human liver slices118	Small (250-μM thick) tissue disc generated from fresh human liver suitable for multiwell plating	Suitable for short-term (~3-day) transcriptomic studies of drug treatments, but direct analysis of NASH tissues is rarely possible.
Organova: bioprinted human liver119	Scaffold-free assembly of human hepatocytes, stellate cells and umbilical vein cells in a 3D in vivo–relevant architecture	Primarily optimized for drug toxicity evaluation, but conditions that mimic NASH are being developed.
Hemoshear: in vitro–engineered liver model117	Primary human hepatocyte, stellate cell and macrophage coculture that incorporates sinusoidal flow in a lipotoxic milieu	Designed primarily to test NASH therapeutics, with transcriptomic and metabolic pathways resembling human disease.
Rat and mouse models
Dietary
Methionine and choline deficiency (MCD)163	High-fat and sucrose diet, deficient in methionine and choline	The histologic appearance in 2–4 weeks resembles human NASH, but animals lose weight and are not insulin resistant, with lack of concordance of the liver transcriptome with human NASH.
Choline-deficient, l-amino acid–defined supplemented, high-fat diet (CDAA)164	High-fat, choline-deficient, reduced-methionine diet for 24 weeks	Like the MCD diet, animals do not gain weight or exhibit insulin resistance, but can develop fibrosis within 6–8 weeks.
High-fat, fructose and cholesterol (AMLN diet)165	Diet high in fat (40%, of which 18% is trans-fat), plus fructose (22%) and cholesterol (2%) administered for 30 weeks	Animals develop both histologic and metabolic features of human NASH with diffuse fibrosis but no cirrhosis.
Genetic
ob/ob166	Leptin-deficient mice	Animals develop severe insulin resistance and steatosis but are resistant to fibrosis without a second insult (e.g., LPS). Unlike this model, humans with NASH have normal or elevated serum leptin.
db/db167	Leptin receptor–deficient mouse	Severe glucose intolerance, obesity and hepatic steatosis; susceptible to fibrosis with a second insult (e.g., MCD or high-fat diet).
Obese (fa/fa) Zucker rat168	Nonfunctional leptin receptor due to mutation	Severe hyperphagia, insulin resistance and obesity on a high-fat diet, with hepatic steatosis but modest fibrosis.
foz/foz169	Truncating mutation in Alstrom syndrome I (ALMS1) protein, a ciliary protein	Severe obesity, insulin resistance, hepatic steatosis and mild fibrosis on a high-fat diet for 300 days, with hypoadiponectinemia. Phenotype may be strain-dependent.
SREBP-1c transgenic170	Hepatocyte-specific overexpression of SREBP-1c	Severe insulin resistance and spontaneous steatohepatitis with oxidant stress at 30 weeks, with some perivenular fibrosis.
MUP-uPA transgenic171	Hepatocyte-specific overexpression of urokinase plasminogen activator	Severe ER stress, hepatic steatosis, mild fibrosis and HCC on high-fat diet (60% of calories), with TNF-α-driven inflammation.
LDLR knockout172	LDL receptor global knockout	Marked steatosis on a diabetogenic diet for 24 weeks, made worse with addition of dietary cholesterol, and association with inflammation, oxidant stress, increased fatty acid production and mild fibrosis.
Phosphatase and tensin homolog (PTEN) knockout173	Alb-Cre-mediated hepatocyte-specific knockout of PTEN	Spontaneous hepatomegaly, steatohepatitis and triglyceride accumulation with lipogenic gene induction at 40 weeks and high penetrance of ademomas, then carcinomas at 74–78 weeks.
Combinations
Streptozotocin plus high fat-diet (STAM) model174	Streptozotocin administered at 2 days, followed by high-fat diet	Steatohepatitis and mild fibrosis with occasional HCCs at 20 weeks. However, animals have type 1 not type 2 diabetes, do not gain weight, and the liver transcriptome is discordant from human NASH.
DIAMOND mouse84	Inbred isogenic strain of C57Bl6/J and S129S1/svlmj fed a high-fat, high-carbohydrate diet	Progressive steatohepatitis, fibrosis then cirrhosis and HCC at 1 year. High transcriptomic concordance to human NASH.
Western diet plus CCl4 addition121	High-fat, high-fructose, high-cholesterol diet with weekly intraperitoneal CCl4 dosing	Progressive steatosis, insulin resistance and fibrosis at 12 weeks, with cirrhosis and frequent HCC at 24 weeks. High transcriptomic concordance to human NASH.
High-fat diet with thermoneutral housing123	Animals housed at thermoneutral (30 °C) conditions, high-fat diet for 8 weeks	Greatly amplified steatohepatitis compared to standard (22 °C) housing, with altered microbiome, increased inflammatory signaling but modest fibrosis.

Commonly used models are listed here with brief descriptions and relevance to human NASH. Rodent models refer to mice unless otherwise specified. This list is not intended to be all-inclusive and does not include other nonrodent animal models (e.g., pigs and nonhuman primates). LPS, lipopolysaccharide.