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A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision‐Cut Liver Slices
Author(s) -
Paish Hannah L.,
Reed Lee H.,
Brown Helen,
Bryan Mark C.,
Govaere Olivier,
Leslie Jack,
Barksby Ben S.,
Garcia Macia Marina,
Watson Abigail,
Xu Xin,
Zaki Marco Y.W.,
Greaves Laura,
Whitehall Julia,
French Jeremy,
White Steven A.,
Manas Derek M.,
Robinson Stuart M.,
Spoletini Gabriele,
Griffiths Clive,
Mann Derek A.,
Borthwick Lee A.,
Drinnan Michael J.,
Mann Jelena,
Oakley Fiona
Publication year - 2019
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.30651
Subject(s) - nintedanib , fibrosis , hydroxyproline , platelet derived growth factor receptor , biology , hepatic fibrosis , extracellular matrix , albumin , growth factor , transforming growth factor , chemistry , microbiology and biotechnology , endocrinology , biochemistry , medicine , receptor , idiopathic pulmonary fibrosis , lung
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 (TGFβ1) and platelet‐derived growth factor (PDGFββ) stimulation ± therapy. Fibrosis was assessed by gene expression, picrosirius red, and α‐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. TGFβ1/PDGFββ 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 TGFβ1/PDGFββ‐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.