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A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision-Cut Liver Slices.

ABSTRACT: Precision cut liver slices (PCLSs) retain the structure and cellular composition of the native liver and represent an improved system to study liver fibrosis compared to two-dimensional mono- or co-cultures. The aim of this study was to develop a bioreactor system to increase the healthy life span of PCLSs and model fibrogenesis. PCLSs were generated from normal rat or human liver, or fibrotic rat liver, and cultured in our bioreactor. PCLS function was quantified by albumin enzyme-linked immunosorbent assay (ELISA). Fibrosis was induced in PCLSs by transforming growth factor beta 1 (TGF?1) and platelet-derived growth factor (PDGF??) stimulation ± therapy. Fibrosis was assessed by gene expression, picrosirius red, and ?-smooth muscle actin staining, hydroxyproline assay, and soluble ELISAs. Bioreactor-cultured PCLSs are viable, maintaining tissue structure, metabolic activity, and stable albumin secretion for up to 6 days under normoxic culture conditions. Conversely, standard static transwell-cultured PCLSs rapidly deteriorate, and albumin secretion is significantly impaired by 48 hours. TGF?1/PDGF?? stimulation of rat or human PCLSs induced fibrogenic gene expression, release of extracellular matrix proteins, activation of hepatic myofibroblasts, and histological fibrosis. Fibrogenesis slowly progresses over 6 days in cultured fibrotic rat PCLSs without exogenous challenge. Activin receptor-like kinase 5 (Alk5) inhibitor (Alk5i), nintedanib, and obeticholic acid therapy limited fibrogenesis in TGF?1/PDGF??-stimulated PCLSs, and Alk5i blunted progression of fibrosis in fibrotic PCLS. Conclusion: We describe a bioreactor technology that maintains functional PCLS cultures for 6 days. Bioreactor-cultured PCLSs can be successfully used to model fibrogenesis and demonstrate efficacy of antifibrotic therapies.

SUBMITTER: Paish HL

PROVIDER: S-EPMC6852483 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision-Cut Liver Slices.

Paish Hannah L HL Reed Lee H LH Brown Helen H Bryan Mark C MC Govaere Olivier O Leslie Jack J Barksby Ben S BS Garcia Macia Marina M Watson Abigail A Xu Xin X Zaki Marco Y W MYW Greaves Laura L Whitehall Julia J French Jeremy J White Steven A SA Manas Derek M DM Robinson Stuart M SM Spoletini Gabriele G Griffiths Clive C Mann Derek A DA Borthwick Lee A LA Drinnan Michael J MJ Mann Jelena J Oakley Fiona F

Hepatology (Baltimore, Md.) 20190528 4

Precision cut liver slices (PCLSs) retain the structure and cellular composition of the native liver and represent an improved system to study liver fibrosis compared to two-dimensional mono- or co-cultures. The aim of this study was to develop a bioreactor system to increase the healthy life span of PCLSs and model fibrogenesis. PCLSs were generated from normal rat or human liver, or fibrotic rat liver, and cultured in our bioreactor. PCLS function was quantified by albumin enzyme-linked immuno ...[more]

PMID: 30963615

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Project description:Studies in mice have shown that PPAR? is an important regulator of lipid metabolism in liver and key transcription factor involved in the adaptive response to fasting. However, much less is known about the role of PPAR? in human liver.Here we set out to study the function of PPAR? in human liver via analysis of whole genome gene regulation in human liver slices treated with the PPAR? agonist Wy14643.Quantitative PCR indicated that PPAR? is well expressed in human liver and human liver slices and that the classical PPAR? targets PLIN2, VLDLR, ANGPTL4, CPT1A and PDK4 are robustly induced by PPAR? activation. Transcriptomics analysis indicated that 617 genes were upregulated and 665 genes were downregulated by PPAR? activation (q value?<?0.05). Many genes induced by PPAR? activation were involved in lipid metabolism (ACSL5, AGPAT9, FADS1, SLC27A4), xenobiotic metabolism (POR, ABCC2, CYP3A5) or the unfolded protein response, whereas most of the downregulated genes were involved in immune-related pathways. Among the most highly repressed genes upon PPAR? activation were several chemokines (e.g. CXCL9-11, CCL8, CX3CL1, CXCL6), interferon ?-induced genes (e.g. IFITM1, IFIT1, IFIT2, IFIT3) and numerous other immune-related genes (e.g. TLR3, NOS2, and LCN2). Comparative analysis of gene regulation by Wy14643 between human liver slices and primary human hepatocytes showed that down-regulation of gene expression by PPAR? is much better captured by liver slices as compared to primary hepatocytes. In particular, PPAR? activation markedly suppressed immunity/inflammation-related genes in human liver slices but not in primary hepatocytes. Finally, several putative new target genes of PPAR? were identified that were commonly induced by PPAR? activation in the two human liver model systems, including TSKU, RHOF, CA12 and VSIG10L.Our paper demonstrates the suitability and superiority of human liver slices over primary hepatocytes for studying the functional role of PPAR? in human liver. Our data underscore the major role of PPAR? in regulation of hepatic lipid and xenobiotic metabolism in human liver and reveal a marked immuno-suppressive/anti-inflammatory effect of PPAR? in human liver slices that may be therapeutically relevant for non-alcoholic fatty liver disease.

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Dataset Information

A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision-Cut Liver Slices.

Publications

A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision-Cut Liver Slices.

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