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Precision-cut liver slices (PCLS) have been widely utilized in various multicellular studies due to liver architecture and intercellular communication remain preserved. In studying antifibrotic drugs using PCLS, the culture method is currently based on a protocol for applications in drug metabolism and toxicity. This published protocol requires the pre-incubation step to incubate PCLS in a drug-free condition. However, this lag period may limit the effect of anti-fibrotic drugs. Therefore, we aimed to study whether a modification of culture protocol provided benefits in studying anti-fibrotic drugs. In this study, PCLS were cultured with LY2109761, the representative anti-fibrotic drug, by either published or modified protocol. Tissue viability and gene/protein expression of fibrosis-related markers of cultured PCLS were assessed. We found that, despite slightly depressed viability (ATP/protein of PCLS was 71-77% of those cultured by the published protocol), spontaneous fibrosis which is a distinctive feature of PCLS was greatly pronounced by the modified culture protocol. As a result, this novel culture protocol demonstrated apparent inhibitory effect of LY2109761 on the gene expression of fibrosis markers, particularly heat shock protein 47 and alpha smooth muscle actin (the inhibition was higher by 44% and 23% when compared to those cultured by published protocol respectively). This modified culture protocol could be a useful alternative for testing anti-fibrotic drugs using PCLS. Besides the differences in tissue viability and spontaneous fibrosis, the modified culture protocol might reduce time and cost of experiment.

Modified culture protocol for testing anti-fibrotic drugs using precision-cut liver slices