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Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening

Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent, progressive disorder and growing public health concern. To address this issue considerable research has been undertaken in pursuit of new NAFLD therapeutics. Development of effective, high-throughput in vitro models is an important asp...

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Autores principales: Cottier, Karissa E., Bhalerao, Devika, Lewis, Candice, Gaffney, Jeannemarie, Heyward, Scott A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517001/
https://www.ncbi.nlm.nih.gov/pubmed/37739978
http://dx.doi.org/10.1038/s41598-023-42785-9
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author Cottier, Karissa E.
Bhalerao, Devika
Lewis, Candice
Gaffney, Jeannemarie
Heyward, Scott A.
author_facet Cottier, Karissa E.
Bhalerao, Devika
Lewis, Candice
Gaffney, Jeannemarie
Heyward, Scott A.
author_sort Cottier, Karissa E.
collection PubMed
description Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent, progressive disorder and growing public health concern. To address this issue considerable research has been undertaken in pursuit of new NAFLD therapeutics. Development of effective, high-throughput in vitro models is an important aspect of drug discovery. Here, a micropatterned hepatocyte co-culture (MPCC) was used to model liver steatosis. The MPCC model (HEPATOPAC(TM)) is comprised of hepatocytes and 3T3-J2 mouse stromal cells plated onto a patterned standard 96-well or 24-well plate, allowing the cultures to be handled and imaged in a standardized multi-well format. These studies employed high content imaging (HCI) analysis to assess lipid content in cultures. HCI analysis of lipid accumulation allows large numbers of samples to be imaged and analyzed in a relatively short period of time compared to manual acquisition and analysis methods. Treatment of MPCC with free fatty acids (FFA), high glucose and fructose (HGF), or a combination of both induces hepatic steatosis. MPCC treatment with ACC1/ACC2 inhibitors, as either a preventative or reversal agent, showed efficacy against FFA induced hepatic steatosis. Drug induced steatosis was also evaluated. Treatment with valproic acid showed steatosis induction in a lean background, which was significantly potentiated in a fatty liver background. Additionally, these media treatments changed expression of fatty liver related genes. Treatment of MPCC with FFA, HGF, or a combination reversibly altered expression of genes involved in fatty acid metabolism, insulin signaling, and lipid transport. Together, these data demonstrate that MPCC is an easy to use, long-term functional in vitro model of NAFLD having utility for compound screening, drug toxicity evaluation, and assessment of gene regulation.
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spelling pubmed-105170012023-09-24 Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening Cottier, Karissa E. Bhalerao, Devika Lewis, Candice Gaffney, Jeannemarie Heyward, Scott A. Sci Rep Article Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent, progressive disorder and growing public health concern. To address this issue considerable research has been undertaken in pursuit of new NAFLD therapeutics. Development of effective, high-throughput in vitro models is an important aspect of drug discovery. Here, a micropatterned hepatocyte co-culture (MPCC) was used to model liver steatosis. The MPCC model (HEPATOPAC(TM)) is comprised of hepatocytes and 3T3-J2 mouse stromal cells plated onto a patterned standard 96-well or 24-well plate, allowing the cultures to be handled and imaged in a standardized multi-well format. These studies employed high content imaging (HCI) analysis to assess lipid content in cultures. HCI analysis of lipid accumulation allows large numbers of samples to be imaged and analyzed in a relatively short period of time compared to manual acquisition and analysis methods. Treatment of MPCC with free fatty acids (FFA), high glucose and fructose (HGF), or a combination of both induces hepatic steatosis. MPCC treatment with ACC1/ACC2 inhibitors, as either a preventative or reversal agent, showed efficacy against FFA induced hepatic steatosis. Drug induced steatosis was also evaluated. Treatment with valproic acid showed steatosis induction in a lean background, which was significantly potentiated in a fatty liver background. Additionally, these media treatments changed expression of fatty liver related genes. Treatment of MPCC with FFA, HGF, or a combination reversibly altered expression of genes involved in fatty acid metabolism, insulin signaling, and lipid transport. Together, these data demonstrate that MPCC is an easy to use, long-term functional in vitro model of NAFLD having utility for compound screening, drug toxicity evaluation, and assessment of gene regulation. Nature Publishing Group UK 2023-09-22 /pmc/articles/PMC10517001/ /pubmed/37739978 http://dx.doi.org/10.1038/s41598-023-42785-9 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Cottier, Karissa E.
Bhalerao, Devika
Lewis, Candice
Gaffney, Jeannemarie
Heyward, Scott A.
Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title_full Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title_fullStr Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title_full_unstemmed Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title_short Micropatterned primary hepatocyte co-culture (HEPATOPAC) for fatty liver disease modeling and drug screening
title_sort micropatterned primary hepatocyte co-culture (hepatopac) for fatty liver disease modeling and drug screening
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517001/
https://www.ncbi.nlm.nih.gov/pubmed/37739978
http://dx.doi.org/10.1038/s41598-023-42785-9
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