A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture

Abstract Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells, but current methods require large numbers of animals which is unwanted in view of the 3R principle and introduces variation. Moreover, stringent breed...

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Autores principales:	Evelien Eenjes, Tinne C. J. Mertens, Marjon J. Buscop-van Kempen, Yolanda van Wijck, Christian Taube, Robbert J. Rottier, Pieter S. Hiemstra
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c736bd1833744ebba2e7b5a8a49cd4ed
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spelling	oai:doaj.org-article:c736bd1833744ebba2e7b5a8a49cd4ed2021-12-02T11:41:24ZA novel method for expansion and differentiation of mouse tracheal epithelial cells in culture10.1038/s41598-018-25799-62045-2322https://doaj.org/article/c736bd1833744ebba2e7b5a8a49cd4ed2018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-25799-6https://doaj.org/toc/2045-2322Abstract Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells, but current methods require large numbers of animals which is unwanted in view of the 3R principle and introduces variation. Moreover, stringent breeding schemes are frequently needed to generate sufficient numbers of genetically modified animals. Current protocols do not incorporate expansion of MTEC, and therefore we developed a protocol to expand MTEC while maintaining their differentiation capacity. MTEC were isolated and expanded using the ROCK inhibitor Y-27632 in presence or absence of the γ-secretase inhibitor DAPT, a Notch pathway inhibitor. Whereas MTEC proliferated without DAPT, growth rate and cell morphology improved in presence of DAPT. ALI-induced differentiation of expanded MTEC resulted in an altered capacity of basal cells to differentiate into ciliated cells, whereas IL-13-induced goblet cell differentiation remained unaffected. Ciliated cell differentiation improved by prolonging the ALI differentiation or by adding DAPT, suggesting that basal cells retain their ability to differentiate. This technique using expansion of MTEC and subsequent ALI differentiation drastically reduces animal numbers and costs for in vitro experiments, and will reduce biological variation. Additionally, we provide novel insights in the dynamics of basal cell populations in vitro.Evelien EenjesTinne C. J. MertensMarjon J. Buscop-van KempenYolanda van WijckChristian TaubeRobbert J. RottierPieter S. HiemstraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Evelien Eenjes Tinne C. J. Mertens Marjon J. Buscop-van Kempen Yolanda van Wijck Christian Taube Robbert J. Rottier Pieter S. Hiemstra A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
description	Abstract Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells, but current methods require large numbers of animals which is unwanted in view of the 3R principle and introduces variation. Moreover, stringent breeding schemes are frequently needed to generate sufficient numbers of genetically modified animals. Current protocols do not incorporate expansion of MTEC, and therefore we developed a protocol to expand MTEC while maintaining their differentiation capacity. MTEC were isolated and expanded using the ROCK inhibitor Y-27632 in presence or absence of the γ-secretase inhibitor DAPT, a Notch pathway inhibitor. Whereas MTEC proliferated without DAPT, growth rate and cell morphology improved in presence of DAPT. ALI-induced differentiation of expanded MTEC resulted in an altered capacity of basal cells to differentiate into ciliated cells, whereas IL-13-induced goblet cell differentiation remained unaffected. Ciliated cell differentiation improved by prolonging the ALI differentiation or by adding DAPT, suggesting that basal cells retain their ability to differentiate. This technique using expansion of MTEC and subsequent ALI differentiation drastically reduces animal numbers and costs for in vitro experiments, and will reduce biological variation. Additionally, we provide novel insights in the dynamics of basal cell populations in vitro.
format	article
author	Evelien Eenjes Tinne C. J. Mertens Marjon J. Buscop-van Kempen Yolanda van Wijck Christian Taube Robbert J. Rottier Pieter S. Hiemstra
author_facet	Evelien Eenjes Tinne C. J. Mertens Marjon J. Buscop-van Kempen Yolanda van Wijck Christian Taube Robbert J. Rottier Pieter S. Hiemstra
author_sort	Evelien Eenjes
title	A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
title_short	A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
title_full	A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
title_fullStr	A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
title_full_unstemmed	A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
title_sort	novel method for expansion and differentiation of mouse tracheal epithelial cells in culture
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/c736bd1833744ebba2e7b5a8a49cd4ed
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_version_	1718395426008203264

A novel method for expansion and differentiation of mouse tracheal epithelial cells in culture

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