Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro

Abstract In vitro human liver models are critical to mitigate species-specific differences observed for toxicology, disease modeling, and regenerative medicine. Interactions with mesenchyme (i.e., fibroblasts) can promote phenotypic functions of primary human hepatocytes (PHHs) in culture; however,...

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Main Authors: Grace E. Brown, Vedant V. Bodke, Brenton R. Ware, Salman R. Khetani
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08135-3
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author Grace E. Brown
Vedant V. Bodke
Brenton R. Ware
Salman R. Khetani
author_facet Grace E. Brown
Vedant V. Bodke
Brenton R. Ware
Salman R. Khetani
author_sort Grace E. Brown
collection DOAJ
description Abstract In vitro human liver models are critical to mitigate species-specific differences observed for toxicology, disease modeling, and regenerative medicine. Interactions with mesenchyme (i.e., fibroblasts) can promote phenotypic functions of primary human hepatocytes (PHHs) in culture; however, using liver-derived fibroblasts remains elusive. Portal fibroblasts (PFs) around the portal triad influence bile duct formation during development, but their role in regulating homeostatic hepatic functions remains unknown. Here, we show that human liver PFs induce long-term phenotypic functions in PHHs at higher levels than activated hepatic stellate cells across 2-dimensional and 3-dimensional culture formats. While PF-conditioned media induces some hepatic functions, partly via insulin-like growth factor binding protein-5 signaling, direct contact is necessary to induce optimal functional levels. Inhibiting Notch signaling reduces progenitor-like characteristics of PHHs and further enhances functionality. Overall, this work demonstrates a unique role for PFs in modulating hepatic functions and provides all-human and all-liver coculture strategies for downstream applications.
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spelling doaj-art-b74b1a15faf247e48df26b126cef49e42025-08-20T03:09:19ZengNature PortfolioCommunications Biology2399-36422025-05-018111710.1038/s42003-025-08135-3Liver portal fibroblasts induce the functions of primary human hepatocytes in vitroGrace E. Brown0Vedant V. Bodke1Brenton R. Ware2Salman R. Khetani3Department of Biomedical Engineering, University of Illinois ChicagoDepartment of Biomedical Engineering, University of Illinois ChicagoDepartment of Biomedical Engineering, University of Illinois ChicagoDepartment of Biomedical Engineering, University of Illinois ChicagoAbstract In vitro human liver models are critical to mitigate species-specific differences observed for toxicology, disease modeling, and regenerative medicine. Interactions with mesenchyme (i.e., fibroblasts) can promote phenotypic functions of primary human hepatocytes (PHHs) in culture; however, using liver-derived fibroblasts remains elusive. Portal fibroblasts (PFs) around the portal triad influence bile duct formation during development, but their role in regulating homeostatic hepatic functions remains unknown. Here, we show that human liver PFs induce long-term phenotypic functions in PHHs at higher levels than activated hepatic stellate cells across 2-dimensional and 3-dimensional culture formats. While PF-conditioned media induces some hepatic functions, partly via insulin-like growth factor binding protein-5 signaling, direct contact is necessary to induce optimal functional levels. Inhibiting Notch signaling reduces progenitor-like characteristics of PHHs and further enhances functionality. Overall, this work demonstrates a unique role for PFs in modulating hepatic functions and provides all-human and all-liver coculture strategies for downstream applications.https://doi.org/10.1038/s42003-025-08135-3
spellingShingle Grace E. Brown
Vedant V. Bodke
Brenton R. Ware
Salman R. Khetani
Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
Communications Biology
title Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
title_full Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
title_fullStr Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
title_full_unstemmed Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
title_short Liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
title_sort liver portal fibroblasts induce the functions of primary human hepatocytes in vitro
url https://doi.org/10.1038/s42003-025-08135-3
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AT salmanrkhetani liverportalfibroblastsinducethefunctionsofprimaryhumanhepatocytesinvitro