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Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture
Primary hepatocytes cultured in vitro are a powerful tool to study the functions of hepatocytes and to evaluate the metabolism and toxicity of new drugs. However, in vitro culture of hepatocytes has proven to be very difficult. Ordinary culture conditions lead to dedifferentiation of hepatocytes, re...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5517567/ https://www.ncbi.nlm.nih.gov/pubmed/28724942 http://dx.doi.org/10.1038/s41598-017-06433-3 |
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author | Guo, Ren Xu, Xinxiu Lu, Yuting Xie, Xin |
author_facet | Guo, Ren Xu, Xinxiu Lu, Yuting Xie, Xin |
author_sort | Guo, Ren |
collection | PubMed |
description | Primary hepatocytes cultured in vitro are a powerful tool to study the functions of hepatocytes and to evaluate the metabolism and toxicity of new drugs. However, in vitro culture of hepatocytes has proven to be very difficult. Ordinary culture conditions lead to dedifferentiation of hepatocytes, resulting in rapid change in cell morphology and significant reduction in specific cell functions. In the current study, we show that hepatocyte dedifferentiation is a rapid process under 21% O(2) conditions. Hepatocytes cultured in 21% O(2) undergo epithelial-to-mesenchymal transition (EMT), obtain fibroblast-like morphology, and show decreased hepatic functions. In contrast, 5% O(2) is very effective in maintaining the epithelial morphology and many functions of the primary hepatocytes cultured in vitro for up to five days. These functions include albumin production, glycogen storage, LDL-uptake and CYP450-mediated drug metabolism. Furthermore, we find that 5% O(2) can relieve the production of reactive oxygen species (ROS) and decrease the level of DNA damage in primary cultured hepatocytes. In addition, we also show that blocking the ERK and GSK-3β pathways can inhibit the dedifferentiation of hepatocytes to a certain extent. Lowering the oxygen tension in cell culture is easily achievable, we believe it could be combined with other methods, such as the use of small molecule cocktails and 3D culture, to maintain proliferation and functions of primary hepatocytes in vitro. |
format | Online Article Text |
id | pubmed-5517567 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-55175672017-07-20 Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture Guo, Ren Xu, Xinxiu Lu, Yuting Xie, Xin Sci Rep Article Primary hepatocytes cultured in vitro are a powerful tool to study the functions of hepatocytes and to evaluate the metabolism and toxicity of new drugs. However, in vitro culture of hepatocytes has proven to be very difficult. Ordinary culture conditions lead to dedifferentiation of hepatocytes, resulting in rapid change in cell morphology and significant reduction in specific cell functions. In the current study, we show that hepatocyte dedifferentiation is a rapid process under 21% O(2) conditions. Hepatocytes cultured in 21% O(2) undergo epithelial-to-mesenchymal transition (EMT), obtain fibroblast-like morphology, and show decreased hepatic functions. In contrast, 5% O(2) is very effective in maintaining the epithelial morphology and many functions of the primary hepatocytes cultured in vitro for up to five days. These functions include albumin production, glycogen storage, LDL-uptake and CYP450-mediated drug metabolism. Furthermore, we find that 5% O(2) can relieve the production of reactive oxygen species (ROS) and decrease the level of DNA damage in primary cultured hepatocytes. In addition, we also show that blocking the ERK and GSK-3β pathways can inhibit the dedifferentiation of hepatocytes to a certain extent. Lowering the oxygen tension in cell culture is easily achievable, we believe it could be combined with other methods, such as the use of small molecule cocktails and 3D culture, to maintain proliferation and functions of primary hepatocytes in vitro. Nature Publishing Group UK 2017-07-19 /pmc/articles/PMC5517567/ /pubmed/28724942 http://dx.doi.org/10.1038/s41598-017-06433-3 Text en © The Author(s) 2017 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Guo, Ren Xu, Xinxiu Lu, Yuting Xie, Xin Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title | Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title_full | Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title_fullStr | Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title_full_unstemmed | Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title_short | Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
title_sort | physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5517567/ https://www.ncbi.nlm.nih.gov/pubmed/28724942 http://dx.doi.org/10.1038/s41598-017-06433-3 |
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