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Fatty Liver Disease Models

NASH Mouse Models

Since 1998, ingenious targeting laboratory has supported metabolic liver disease research with custom mouse models enabling mechanistic studies of hepatic steatosis, steatohepatitis, and fibrosis progression underlying the global epidemic of metabolic dysfunction associated fatty liver disease.

NASH MASH mouse models provide essential platforms for investigating the molecular pathways driving progression from simple steatosis to steatohepatitis and cirrhosis, testing hypotheses about lipotoxicity and inflammation, and developing therapies for this increasingly prevalent condition.

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Understanding NASH and MASH

The terminology for fatty liver disease has evolved to better reflect its metabolic etiology:

Current Nomenclature

MAFLD

Metabolic Associated Fatty Liver Disease

Broader term encompassing hepatic steatosis associated with metabolic dysfunction, regardless of alcohol consumption.

MASLD

Metabolic Dysfunction Associated Steatotic Liver Disease

Current preferred terminology for non alcoholic fatty liver disease.

MASH

Metabolic Dysfunction Associated Steatohepatitis

The progressive form with inflammation and hepatocyte injury, previously termed NASH (non alcoholic steatohepatitis).

Disease Progression

1

Simple Steatosis

Hepatic fat accumulation (>5% of hepatocytes) without significant inflammation. Often reversible with lifestyle intervention.

2

Steatohepatitis (MASH)

Steatosis plus lobular inflammation, hepatocyte ballooning, and injury. Risk of progression to fibrosis.

3

Fibrosis

Collagen deposition and architectural distortion. Staged F0 to F4.

4

Cirrhosis

Advanced fibrosis with nodule formation. Risk of hepatocellular carcinoma and liver failure.

Dietary Models

High Fat Diet Models

Standard High Fat Diet (HFD)

45% to 60% fat calories produces obesity and hepatic steatosis in C57BL/6 mice over 12 to 24 weeks. Produces mild steatohepatitis but minimal fibrosis.

Western Diet

High fat plus high fructose/sucrose mimics Western dietary patterns. More severe metabolic phenotype than HFD alone.

Fast Food Diet

High fat, high cholesterol, high fructose combination produces more severe disease with fibrosis.

Specialized Dietary Models

Choline Deficient High Fat Diet (CDHFD)

Choline deficiency accelerates steatohepatitis and fibrosis development. Widely used for rapid disease induction.

Methionine Choline Deficient Diet (MCD)

Produces severe steatohepatitis and fibrosis but with weight loss rather than obesity. Does not model metabolic syndrome.

AMLN Diet (Amylin Liver NASH)

High trans fat, high fructose, high cholesterol diet produces obesity, steatohepatitis, and fibrosis with metabolic syndrome features.

DIAMOND Model

Isogenic C57BL/6J 129S1/SvImJ hybrid on Western diet develops full disease spectrum including hepatocellular carcinoma.

Genetic Models

Lipid Metabolism Genes

PNPLA3 I148M Knockin

The most common genetic risk variant for fatty liver disease in humans. Knockin mice with the human I148M variant develop more severe steatosis and fibrosis on lipogenic diets.

TM6SF2 E167K Knockin

Second most common variant. Affects VLDL secretion and hepatic lipid accumulation.

HSD17B13 Loss of Function

Protective variant in humans. HSD17B13 knockout mice show altered lipid metabolism.

MBOAT7 Variants

Membrane bound O acyltransferase 7 variants affect phospholipid remodeling.

Insulin Signaling and Metabolism

Liver Specific Insulin Receptor Knockout (LIRKO)

Hepatocyte insulin resistance model. Develops steatosis and glucose intolerance.

PTEN Liver Knockout

Severe steatohepatitis and hepatocellular carcinoma development.

FXR Knockout

Farnesoid X receptor deficiency affects bile acid and lipid metabolism.

Inflammatory Pathway Models

NLRP3 Inflammasome

Inflammasome activation drives steatohepatitis. NLRP3 knockout or knockin models for pathway analysis.

ASK1 Pathway

Apoptosis signal regulating kinase 1 mediates lipotoxic injury.

TLR4 and Innate Immunity

Toll like receptor signaling contributes to inflammation and fibrosis.

Model Design Considerations

Background Strain Selection

Metabolic phenotypes are profoundly strain dependent:

C57BL/6

Susceptible to diet induced obesity and steatosis. Standard background for MASH studies.

C57BL/6J vs C57BL/6N

Substrains differ in metabolic phenotypes. C57BL/6J carries Nnt mutation affecting glucose metabolism.

129 Strains

Generally resistant to diet induced obesity but susceptible when combined with genetic modifications.

BALB/c

Different metabolic response than C57BL/6.

Cell Type Specific Approaches

MASH involves multiple liver cell types:

Cell TypeCre DriverApplication
Hepatocyte SpecificAlbumin CreTargets hepatocytes for studying cell autonomous lipid metabolism and injury responses.
Kupffer Cell/MacrophageLysM Cre or CX3CR1 CreTargets resident and infiltrating macrophages for inflammation studies.
Hepatic Stellate CellLRAT CreTargets stellate cells for fibrosis mechanism studies.
CholangiocyteCK19 Cre or Sox9 CreTargets bile duct contributions.

Phenotyping MASH Models

Metabolic Assessment

  • Body Composition:Weight, fat mass, lean mass by DEXA or MRI.
  • Glucose Homeostasis:IPGTT, ITT, fasting glucose and insulin, HOMA IR.
  • Lipid Profiles:Plasma triglycerides, cholesterol, free fatty acids.

Liver Assessment

  • Liver Weight and Triglycerides:Hepatomegaly and hepatic lipid content.
  • Histopathology:H&E for steatosis, inflammation, and ballooning. NAFLD Activity Score (NAS) for standardized grading.
  • Fibrosis Staging:Sirius red or trichrome staining. Alpha SMA immunohistochemistry. Hydroxyproline content.

Therapeutic Applications

MASH models support preclinical development of multiple therapeutic approaches:

FXR agonists
ACC inhibitors
GLP1 analogs
Thyroid hormone receptor agonists
Anti inflammatory approaches
Anti fibrotic therapies

Combination Approaches

Advanced MASH may require combination therapy targeting multiple disease mechanisms. Patient variant knockins, cell type specific deletions, and reporter integration enable sophisticated preclinical studies of complex therapeutic strategies.

Selected Publications in MASH Research

According to PubMed, recent publications demonstrate the utility of genetically engineered mouse models in MASH research:

What Researchers Say

I've been working with iTL over the past 5 years in the production of 3 different genetically altered mice. Not only did iTL help in the design of the mice, but the entire process was transparent with the opportunity at any time along the way to discuss my questions or concerns with scientists who had significant insight into the process. The mice were delivered on time, as billed!

Raghu Mirmira, MD, PhD

University of Chicago

Start Your MASH Model Project

Our scientific consultants are ready to discuss your metabolic liver disease research requirements and recommend the optimal model design for your program. Initial consultation is provided at no charge and includes target analysis, model strategy recommendations, and timeline estimates.

✦ New for 2026

Breeding Scheme Architect

Plan complex multi-allele breeding strategies, calculate expected genotype ratios, and estimate time to experimental cohorts—all before starting your project.

Visualize multi-generation breeding paths
Calculate Mendelian ratios automatically
Estimate timeline to study-ready cohorts

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Allele 1Gene-flox (conditional)
Allele 2Cre-driver (tissue-specific)
TargetHomozygous knockout

→ 3 generations to target genotype

Frequently Asked Questions

Common dietary models include high-fat diet (HFD), methionine-choline-deficient (MCD) diet, Western diet, and choline-deficient amino-acid defined (CDAA) diet. MCD diet rapidly induces steatohepatitis without obesity, while HFD models metabolic syndrome context. Selection depends on research question and timeline.

Disease progression timelines vary by model and dietary protocol. Steatosis develops first, followed by steatohepatitis, then fibrosis with longer challenge periods or accelerating approaches. Genetic modifications can accelerate specific stages: combining conditional knockouts with dietary challenge enables study of mechanisms driving progression from steatosis to fibrosis. Contact us to discuss model selection for your study timeline.

Hepatocyte-specific deletion uses Albumin-Cre for lipid metabolism studies. Macrophage/Kupffer cell deletion uses LysM-Cre or CX3CR1-Cre for inflammation studies. Hepatic stellate cell deletion uses LRAT-Cre for fibrosis mechanism studies. Cholangiocyte targeting uses CK19-Cre or Sox9-Cre.

C57BL/6 is the standard background for MASH studies due to susceptibility to diet-induced obesity and steatosis. Substrains differ: C57BL/6J carries Nnt mutation affecting glucose metabolism. C57BL/6N is commonly used and well-characterized. Both work, but consistency within a study is critical.

Custom model generation includes targeting construct design, ES cell targeting, chimera generation, and germline transmission. Conditional approaches for cell-type-specific studies follow similar workflows. Pre-germline characterization provides early validation of targeting before mouse generation. Contact us for current timeline estimates.

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