Cargando…
Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease
Liver biology and function, drug-induced liver injury (DILI) and liver diseases are difficult to study using current in vitro models such as primary human hepatocyte (PHH) monolayer cultures, as their rapid de-differentiation restricts their usefulness substantially. Thus, we have developed and exte...
Autores principales: | , , , , , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855186/ https://www.ncbi.nlm.nih.gov/pubmed/27143246 http://dx.doi.org/10.1038/srep25187 |
_version_ | 1782430327728242688 |
---|---|
author | Bell, Catherine C. Hendriks, Delilah F. G. Moro, Sabrina M. L. Ellis, Ewa Walsh, Joanne Renblom, Anna Fredriksson Puigvert, Lisa Dankers, Anita C. A. Jacobs, Frank Snoeys, Jan Sison-Young, Rowena L. Jenkins, Rosalind E. Nordling, Åsa Mkrtchian, Souren Park, B. Kevin Kitteringham, Neil R. Goldring, Christopher E. P. Lauschke, Volker M. Ingelman-Sundberg, Magnus |
author_facet | Bell, Catherine C. Hendriks, Delilah F. G. Moro, Sabrina M. L. Ellis, Ewa Walsh, Joanne Renblom, Anna Fredriksson Puigvert, Lisa Dankers, Anita C. A. Jacobs, Frank Snoeys, Jan Sison-Young, Rowena L. Jenkins, Rosalind E. Nordling, Åsa Mkrtchian, Souren Park, B. Kevin Kitteringham, Neil R. Goldring, Christopher E. P. Lauschke, Volker M. Ingelman-Sundberg, Magnus |
author_sort | Bell, Catherine C. |
collection | PubMed |
description | Liver biology and function, drug-induced liver injury (DILI) and liver diseases are difficult to study using current in vitro models such as primary human hepatocyte (PHH) monolayer cultures, as their rapid de-differentiation restricts their usefulness substantially. Thus, we have developed and extensively characterized an easily scalable 3D PHH spheroid system in chemically-defined, serum-free conditions. Using whole proteome analyses, we found that PHH spheroids cultured this way were similar to the liver in vivo and even retained their inter-individual variability. Furthermore, PHH spheroids remained phenotypically stable and retained morphology, viability, and hepatocyte-specific functions for culture periods of at least 5 weeks. We show that under chronic exposure, the sensitivity of the hepatocytes drastically increased and toxicity of a set of hepatotoxins was detected at clinically relevant concentrations. An interesting example was the chronic toxicity of fialuridine for which hepatotoxicity was mimicked after repeated-dosing in the PHH spheroid model, not possible to detect using previous in vitro systems. Additionally, we provide proof-of-principle that PHH spheroids can reflect liver pathologies such as cholestasis, steatosis and viral hepatitis. Combined, our results demonstrate that the PHH spheroid system presented here constitutes a versatile and promising in vitro system to study liver function, liver diseases, drug targets and long-term DILI. |
format | Online Article Text |
id | pubmed-4855186 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-48551862016-05-18 Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease Bell, Catherine C. Hendriks, Delilah F. G. Moro, Sabrina M. L. Ellis, Ewa Walsh, Joanne Renblom, Anna Fredriksson Puigvert, Lisa Dankers, Anita C. A. Jacobs, Frank Snoeys, Jan Sison-Young, Rowena L. Jenkins, Rosalind E. Nordling, Åsa Mkrtchian, Souren Park, B. Kevin Kitteringham, Neil R. Goldring, Christopher E. P. Lauschke, Volker M. Ingelman-Sundberg, Magnus Sci Rep Article Liver biology and function, drug-induced liver injury (DILI) and liver diseases are difficult to study using current in vitro models such as primary human hepatocyte (PHH) monolayer cultures, as their rapid de-differentiation restricts their usefulness substantially. Thus, we have developed and extensively characterized an easily scalable 3D PHH spheroid system in chemically-defined, serum-free conditions. Using whole proteome analyses, we found that PHH spheroids cultured this way were similar to the liver in vivo and even retained their inter-individual variability. Furthermore, PHH spheroids remained phenotypically stable and retained morphology, viability, and hepatocyte-specific functions for culture periods of at least 5 weeks. We show that under chronic exposure, the sensitivity of the hepatocytes drastically increased and toxicity of a set of hepatotoxins was detected at clinically relevant concentrations. An interesting example was the chronic toxicity of fialuridine for which hepatotoxicity was mimicked after repeated-dosing in the PHH spheroid model, not possible to detect using previous in vitro systems. Additionally, we provide proof-of-principle that PHH spheroids can reflect liver pathologies such as cholestasis, steatosis and viral hepatitis. Combined, our results demonstrate that the PHH spheroid system presented here constitutes a versatile and promising in vitro system to study liver function, liver diseases, drug targets and long-term DILI. Nature Publishing Group 2016-05-04 /pmc/articles/PMC4855186/ /pubmed/27143246 http://dx.doi.org/10.1038/srep25187 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Bell, Catherine C. Hendriks, Delilah F. G. Moro, Sabrina M. L. Ellis, Ewa Walsh, Joanne Renblom, Anna Fredriksson Puigvert, Lisa Dankers, Anita C. A. Jacobs, Frank Snoeys, Jan Sison-Young, Rowena L. Jenkins, Rosalind E. Nordling, Åsa Mkrtchian, Souren Park, B. Kevin Kitteringham, Neil R. Goldring, Christopher E. P. Lauschke, Volker M. Ingelman-Sundberg, Magnus Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title | Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title_full | Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title_fullStr | Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title_full_unstemmed | Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title_short | Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
title_sort | characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855186/ https://www.ncbi.nlm.nih.gov/pubmed/27143246 http://dx.doi.org/10.1038/srep25187 |
work_keys_str_mv | AT bellcatherinec characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT hendriksdelilahfg characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT morosabrinaml characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT ellisewa characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT walshjoanne characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT renblomanna characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT fredrikssonpuigvertlisa characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT dankersanitaca characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT jacobsfrank characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT snoeysjan characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT sisonyoungrowenal characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT jenkinsrosalinde characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT nordlingasa characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT mkrtchiansouren characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT parkbkevin characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT kitteringhamneilr characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT goldringchristopherep characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT lauschkevolkerm characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease AT ingelmansundbergmagnus characterizationofprimaryhumanhepatocytespheroidsasamodelsystemfordruginducedliverinjuryliverfunctionanddisease |