Developing an in vitro screening assay platform for evaluation of antifibrotic drugs using precision-cut liver slices

• DOI: 10.1186/s13069-014-0017-2
• Publication Date: 2015-01-09T12:29:17Z
• Authors: Satish Kumar Sada...; Nethra Siddaraju; Khaiser Mehdi Khan; Balamuralikrishna...; Nimisha R Kumar; Vibha Haridas; Madhusudhan B Reddy; Somesh Baggavalli; Anup M Oommen; Raghavendra Pralh...
• Keywords: Hepatic stellate cell; Steatohepatitis; Myofibroblast; Lumican; Steatosis

Precision-cut liver slices present different cell types of in a physiological context, and they have been explored as effective vitro model systems to study fibrosis. Inducing fibrosis the using toxicants like carbon tetrachloride is less relevance human disease conditions. Our aim for this was establish physiologically relevant conditions induce fibrotic phenotypes slices. 150 μm thickness were obtained from female C57BL/6 J mice. The cultured 24 hours media containing cocktail 10 nM each TGF-β, PDGF, 5 μM lysophosphatidic acid sphingosine 1 phosphate 0.2 μg/ml lipopolysaccharide along with 500 palmitate analyzed triglyceride accumulation, stress inflammation, myofibroblast activation extracellular matrix (ECM) accumulation. Incubation resulted increased hallmark steatosis. levels Acta2, inflammatory genes (IL-6, TNF-α C-reactive protein) significantly elevated. In addition, treatment ECM markers - collagen, lumican fibronectin. This reports experimental required associated steatohepatitis inducers. system presented here captures various aspects process steatosis, stellate accumulation serves platform screen small molecules their antifibrotic activity.