Precision‐cut liver slice culture (PCLSC) ex vivo model of steatohepatitis

Lipotoxicity caused by alcohol misuse or obesity mediates insulin resistance and progressive liver disease, but the mechanisms are poorly understood. PCLSCs provide 3‐D ex vivo models in which all tissue cell types are represented with normal architectural relationships. We used 150 μm thick adult rat PCLSCs to study mediators of lipotoxicity, focusing on the roles of long‐chain saturated versus unsaturated fatty acids and ceramides. Cultures were exposed for 48h to vehicle, or 0.5 mM palmitic (C16:0‐saturated; PA) or oleic (C18:1‐mono‐unsaturated OA) acid ± myriocin to inhibit ceramide synthesis. Both PA and OA increased G6PD release and OA increased hepatic neutral lipids. PA exposure caused hepatic steatosis with increased clusters of oval and stellate cells. OA caused striking micro‐vesicular steatohepatitis with necrosis and apoptosis. Myriocin nearly prevented hepatic steatosis, steatohepatitis, and oval/stellate cell proliferation in PA and OA treated cultures. These results illustrate the utility of the PCLSC model for mechanistic studies of non‐alcoholic steatohepatitis, and further demonstrate the role of toxic ceramides as mediators of lipotoxicity and hepatocellular injury. Research supported by AA‐11431, AA‐12908, and AA‐16126 from the National Institutes of Health