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Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors
Most in vitro test systems for the assessment of toxicity are based on endpoint measurements and cannot contribute much to the establishment of mechanistic models, which are crucially important for further progress in this field. Hence, in recent years, much effort has been put into the development...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7426412/ https://www.ncbi.nlm.nih.gov/pubmed/32792676 http://dx.doi.org/10.1038/s41598-020-70785-6 |
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author | Gehre, Christian Flechner, Marie Kammerer, Sarah Küpper, Jan-Heiner Coleman, Charles Dominic Püschel, Gerhard Paul Uhlig, Katja Duschl, Claus |
author_facet | Gehre, Christian Flechner, Marie Kammerer, Sarah Küpper, Jan-Heiner Coleman, Charles Dominic Püschel, Gerhard Paul Uhlig, Katja Duschl, Claus |
author_sort | Gehre, Christian |
collection | PubMed |
description | Most in vitro test systems for the assessment of toxicity are based on endpoint measurements and cannot contribute much to the establishment of mechanistic models, which are crucially important for further progress in this field. Hence, in recent years, much effort has been put into the development of methods that generate kinetic data. Real time measurements of the metabolic activity of cells based on the use of oxygen sensitive microsensor beads have been shown to provide access to the mode of action of compounds in hepatocytes. However, for fully exploiting this approach a detailed knowledge of the microenvironment of the cells is required. In this work, we investigate the cellular behaviour of three types of hepatocytes, HepG2 cells, HepG2-3A4 cells and primary mouse hepatocytes, towards their exposure to acetaminophen when the availability of oxygen for the cell is systematically varied. We show that the relative emergence of two modes of action, one NAPQI dependent and the other one transient and NAPQI independent, scale with expression level of CYP3A4. The transient cellular response associated to mitochondrial respiration is used to characterise the influence of the initial oxygen concentration in the wells before exposure to acetaminophen on the cell behaviour. A simple model is presented to describe the behaviour of the cells in this scenario. It demonstrates the level of control over the role of oxygen supply in these experiments. This is crucial for establishing this approach into a reliable and powerful method for the assessment of toxicity. |
format | Online Article Text |
id | pubmed-7426412 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-74264122020-08-14 Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors Gehre, Christian Flechner, Marie Kammerer, Sarah Küpper, Jan-Heiner Coleman, Charles Dominic Püschel, Gerhard Paul Uhlig, Katja Duschl, Claus Sci Rep Article Most in vitro test systems for the assessment of toxicity are based on endpoint measurements and cannot contribute much to the establishment of mechanistic models, which are crucially important for further progress in this field. Hence, in recent years, much effort has been put into the development of methods that generate kinetic data. Real time measurements of the metabolic activity of cells based on the use of oxygen sensitive microsensor beads have been shown to provide access to the mode of action of compounds in hepatocytes. However, for fully exploiting this approach a detailed knowledge of the microenvironment of the cells is required. In this work, we investigate the cellular behaviour of three types of hepatocytes, HepG2 cells, HepG2-3A4 cells and primary mouse hepatocytes, towards their exposure to acetaminophen when the availability of oxygen for the cell is systematically varied. We show that the relative emergence of two modes of action, one NAPQI dependent and the other one transient and NAPQI independent, scale with expression level of CYP3A4. The transient cellular response associated to mitochondrial respiration is used to characterise the influence of the initial oxygen concentration in the wells before exposure to acetaminophen on the cell behaviour. A simple model is presented to describe the behaviour of the cells in this scenario. It demonstrates the level of control over the role of oxygen supply in these experiments. This is crucial for establishing this approach into a reliable and powerful method for the assessment of toxicity. Nature Publishing Group UK 2020-08-13 /pmc/articles/PMC7426412/ /pubmed/32792676 http://dx.doi.org/10.1038/s41598-020-70785-6 Text en © The Author(s) 2020 Open AccessThis 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/. |
spellingShingle | Article Gehre, Christian Flechner, Marie Kammerer, Sarah Küpper, Jan-Heiner Coleman, Charles Dominic Püschel, Gerhard Paul Uhlig, Katja Duschl, Claus Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title | Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title_full | Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title_fullStr | Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title_full_unstemmed | Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title_short | Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
title_sort | real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7426412/ https://www.ncbi.nlm.nih.gov/pubmed/32792676 http://dx.doi.org/10.1038/s41598-020-70785-6 |
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