Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture

Abstract 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 hepato...

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Autores principales:	Ren Guo, Xinxiu Xu, Yuting Lu, Xin Xie
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0607e8f74a4f4a73b8c44b71bb6a4618
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spelling	oai:doaj.org-article:0607e8f74a4f4a73b8c44b71bb6a46182021-12-02T12:32:51ZPhysiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture10.1038/s41598-017-06433-32045-2322https://doaj.org/article/0607e8f74a4f4a73b8c44b71bb6a46182017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06433-3https://doaj.org/toc/2045-2322Abstract 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% O2 conditions. Hepatocytes cultured in 21% O2 undergo epithelial-to-mesenchymal transition (EMT), obtain fibroblast-like morphology, and show decreased hepatic functions. In contrast, 5% O2 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% O2 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.Ren GuoXinxiu XuYuting LuXin XieNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ren Guo Xinxiu Xu Yuting Lu Xin Xie Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture
description	Abstract 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% O2 conditions. Hepatocytes cultured in 21% O2 undergo epithelial-to-mesenchymal transition (EMT), obtain fibroblast-like morphology, and show decreased hepatic functions. In contrast, 5% O2 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% O2 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	article
author	Ren Guo Xinxiu Xu Yuting Lu Xin Xie
author_facet	Ren Guo Xinxiu Xu Yuting Lu Xin Xie
author_sort	Ren Guo
title	Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture
title_short	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_sort	physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/0607e8f74a4f4a73b8c44b71bb6a4618
work_keys_str_mv	AT renguo physiologicaloxygentensionreduceshepatocytededifferentiationininvitroculture AT xinxiuxu physiologicaloxygentensionreduceshepatocytededifferentiationininvitroculture AT yutinglu physiologicaloxygentensionreduceshepatocytededifferentiationininvitroculture AT xinxie physiologicaloxygentensionreduceshepatocytededifferentiationininvitroculture
_version_	1718393898833805312

Physiological oxygen tension reduces hepatocyte dedifferentiation in in vitro culture

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