How to accurately model the complexities of human NASH in vitro?
A common cause of chronic liver disease worldwide is Non-Alcoholic Fatty Liver Disease (NAFLD) – a spectrum of metabolic diseases where fat builds up in the liver in the absence of excessive alcohol consumption. 20% of people diagnosed with NAFLD develop NASH. Left untreated, NASH may progress into cirrhosis and/or liver cancer.
Despite its prevalence, this disease remains unmet therapeutically; why is this? NAFLD/NASH arises from multiple factors that result in phenotypes that are difficult to replicate in the laboratory. The inability of current preclinical models to replicate human NASH is a contributing factor to drug attrition.
The PhysioMimix® NASH Assay utilizes a tri-culture Liver-on-a-chip model comprised of human primary liver cells grown in 3D, under perfusion to recapitulate the liver’s physiological environment.
Primary human hepatocytes, Kupffer cells (inflammatory response), and stellate cells (fibrotic response) cultures are treated with proprietary HEP-Fat media to create an in vitro NASH phenotype that closely mimics the key aspects of the disease.
This advanced assay delivers long-term 3D liver microtissue cultures that enable the assessment of therapeutic efficacy or the exploration of disease pathways for target identification. We utilize validated cells with known genotypes, allowing NASH-related single nucleotide polymorphisms (SNPs) to be explored.
The NASH assay is functional for at least 14 days. It enables the quantification of biomarkers (such as inflammation, fibrosis, and steatosis) and cell health markers (including ALT and AST), allowing translatability to the clinic.
Limitations of current techniques
- Lack of human-relevant preclinical models that capture disease complexity
- Animal models failing to predict clinical outcomes
- Poor translatability of preclinical data
- Short-lived hepatic function in standard cultures limits experiments
- Difficulty tailoring models to suit specific scientific questions
Advancements with PhysioMimix OOC®
- PhysioMimix NASH model replicates key phenotypic biomarkers
- Generate data using key hepatic human cell types to support in vivo findings
- Measure key clinical markers such as ALT and AST
- Phenotypic NASH cultures for at least 14 days to enable a longer-term analysis
- Highly adaptable to specific aspects of the disease to tailor the application to suit your needs
View our “Pros and cons of preclinical models of non-alcoholic steatohepatitis” infographic
Standard NASH cell culture timeline
End point measurements
Longitudinal and endpoint measurements include (but not limited to):
- Albumin production
- Urea production
Clinical liver heath biomarkers
- Lactose dehydrogenase (LDH) release
- Aspartate Transferase (AST)/Alanine amino transferase (ALT)
- Luminex®/ELISA assays
- Fibrosis (e.g. TIMP-1, Pro-collagen, Fibronectin)
- Inflammation (e.g. IL-6, IL-8 TNF-α)
- Confocal microscopy
- Smooth muscle actin
- Fat accumulation (Nile Red staining)
Optional profiling analysis
- Quantitative PCR
Recapitulating key aspects of human NASH disease
Our PhysioMimix NASH model captures key aspects of human disease including steatosis, inflammation, and fibrosis. By assessing biomarker expression, the assay provides high-content quantitative data from every replicate, including an assessment of fibrosis.
Improved alignment of transcriptomic profiles to human
Transcriptomic profiling of the PhysioMimix NASH model demonstrates a strong NASH profile when compared to human data and more closely replicates changes found in NASH patients than the murine WD model (Vacca et al., 2020).
Access our NAFLD/NASH Service
Get instant access to the PhysioMimix NAFLD/NASH Assay via our CRO Service. Through a collaborative approach, our experts work with you to plan and execute your study.
Standard and bespoke projects are carried out by our dedicated team of scientists in our CRO facility providing you with actionable data within weeks.
PhysioMimix NASH-in-a-box kit
NASH-in-a-box contains everything that you require to recreate our PhysioMimix NASH Assay in your own laboratory.
Simply follow the software-guided protocol on your PhysioMimix Single- or Multi-organ microphysiological system to produce clinically translatable data fast.