Generation of proliferating human adult hepatocytes using optimized 3D culture conditions

Abstract Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we described optimized 3D culture condi...

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Autores principales:	Sophie Rose, Frédéric Ezan, Marie Cuvellier, Arnaud Bruyère, Vincent Legagneux, Sophie Langouët, Georges Baffet
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5698f8c84c2c4af488edf5e3538a8768
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spelling	oai:doaj.org-article:5698f8c84c2c4af488edf5e3538a87682021-12-02T14:12:45ZGeneration of proliferating human adult hepatocytes using optimized 3D culture conditions10.1038/s41598-020-80019-42045-2322https://doaj.org/article/5698f8c84c2c4af488edf5e3538a87682021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80019-4https://doaj.org/toc/2045-2322Abstract Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we described optimized 3D culture conditions of primary human hepatocytes (PHH) to trigger two waves of proliferation and we identified matrix stiffness and cell–cell interactions as the main actors necessary for this proliferation. We demonstrated that DNA replication and overexpression of cell cycle markers are modulate by the matrix stiffness while PHH cultured in 3D without prior cellular interactions did not proliferate. Besides, we showed that PHH carry out an additional cell cycle after transient inhibition of MAPK MER1/2-ERK1/2 signaling pathway. Collagen cultured hepatocytes are organized as characteristic hollow spheroids able to maintain survival, cell polarity and hepatic differentiation for long-term culture periods of at least 28 days. Remarkably, we demonstrated by transcriptomic analysis and functional experiments that proliferating cells are mature hepatocytes with high detoxication capacities. In conclusion, the advanced 3D model described here, named Hepoid, is particularly relevant for obtaining normal human proliferating hepatocytes. By allowing concomitant proliferation and differentiation, it constitutes a promising tool for many pharmacological and biotechnological applications.Sophie RoseFrédéric EzanMarie CuvellierArnaud BruyèreVincent LegagneuxSophie LangouëtGeorges BaffetNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sophie Rose Frédéric Ezan Marie Cuvellier Arnaud Bruyère Vincent Legagneux Sophie Langouët Georges Baffet Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
description	Abstract Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we described optimized 3D culture conditions of primary human hepatocytes (PHH) to trigger two waves of proliferation and we identified matrix stiffness and cell–cell interactions as the main actors necessary for this proliferation. We demonstrated that DNA replication and overexpression of cell cycle markers are modulate by the matrix stiffness while PHH cultured in 3D without prior cellular interactions did not proliferate. Besides, we showed that PHH carry out an additional cell cycle after transient inhibition of MAPK MER1/2-ERK1/2 signaling pathway. Collagen cultured hepatocytes are organized as characteristic hollow spheroids able to maintain survival, cell polarity and hepatic differentiation for long-term culture periods of at least 28 days. Remarkably, we demonstrated by transcriptomic analysis and functional experiments that proliferating cells are mature hepatocytes with high detoxication capacities. In conclusion, the advanced 3D model described here, named Hepoid, is particularly relevant for obtaining normal human proliferating hepatocytes. By allowing concomitant proliferation and differentiation, it constitutes a promising tool for many pharmacological and biotechnological applications.
format	article
author	Sophie Rose Frédéric Ezan Marie Cuvellier Arnaud Bruyère Vincent Legagneux Sophie Langouët Georges Baffet
author_facet	Sophie Rose Frédéric Ezan Marie Cuvellier Arnaud Bruyère Vincent Legagneux Sophie Langouët Georges Baffet
author_sort	Sophie Rose
title	Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
title_short	Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
title_full	Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
title_fullStr	Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
title_full_unstemmed	Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
title_sort	generation of proliferating human adult hepatocytes using optimized 3d culture conditions
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/5698f8c84c2c4af488edf5e3538a8768
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Generation of proliferating human adult hepatocytes using optimized 3D culture conditions

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