Non-alcoholic steatohepatitis (NASH) is a progressive disease which results in advancement of liver fibrosis, cirrhosis, and hepatocellular carcinoma, for which there are no FDA-approved drug therapies. Nearly half of the patients suffering from NAFLD can progress to non-alcoholic steatohepatitis (NASH), an inflammatory syndrome which disrupts the liver’s function. Thus, it is important to design various drug discovery assays for this problem.
Visikol is one of the leaders in developing various in vitro assays and 3D cell culture models. The quantitative evaluation of tissue imaging services as well as cell-based assays are valuable and at Visikol, the uniquely designed assays with advanced cell culture techniques and high content imaging help to enable quantitative evaluation of promising compounds in vitro.
Visikol HepaRG/NP 3D spheroids were pretreated with either vehicle or an ALK-5 inhibitor for 1 h prior to inducing fibrosis with 100 nM TGF-β for 48 h. DAPI in blue, viability indicator in green, pan collagen in red.
NASH Co-Culture Model
Visikol has developed a co-culture model for NASH which is very useful in studying the pathogenesis of the NASH disease and screening of drug compounds. To develop a human liver spheroid/NASH model, a hetero-cellular spheroid system comprised of primary human hepatocytes (PHH) are co-cultured with fractions of primary human liver non-parenchymal cells (NPC) into ULA U-bottom plates and maintained under standard culture conditions for 14 days to enable spheroid aggregation to a size of approximately 200 μm in diameter. After the formation of spheroids, they are pre-treated (other timepoints available upon request) with test articles and fibrosis is induced via addition of TGF-β for an additional 72 hours. Test article concentration is maintained through induction of fibrosis. For NASH like phenotype, spheroids are induced with FFA (palmitic acid and Oleic acid) from day 5 to day 14 of the spheroid formation. Then the spheroids are collected and pooled and collected for various endpoints such as RNA extraction, immunofluorescent labeling, high content imaging and ELISAs for protein of interest. Supernatants are collected and stored at -80 C.
Various multicellular human disease models such as NASH are now successful in recapitulating complex in vivo conditions. They are very well suited for mechanistic approach for the disease progression and drug efficacy screening. These models are also helpful in identifying the compounds to reduce biomarkers as there are no approved treatments available for NASH.