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Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization

Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. Although many efforts have been made to reveal the underlying mechanisms of decidualization, the exact signaling between the epithelial cells tha...

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Autores principales: De Clercq, Katrien, Hennes, Aurélie, Vriens, Joris
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408775/
https://www.ncbi.nlm.nih.gov/pubmed/28287563
http://dx.doi.org/10.3791/55168
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author De Clercq, Katrien
Hennes, Aurélie
Vriens, Joris
author_facet De Clercq, Katrien
Hennes, Aurélie
Vriens, Joris
author_sort De Clercq, Katrien
collection PubMed
description Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. Although many efforts have been made to reveal the underlying mechanisms of decidualization, the exact signaling between the epithelial cells that are in contact with the embryo and the underlying stromal cells remains poorly understood. Therefore, studying decidualization in a way that takes both the epithelial and stromal cells into account could improve our knowledge about the molecular details of decidualization. For this purpose, in vivo models of artificial decidualization are physiologically the most relevant; however, manipulation of intercellular communication is limited. Currently, in vitro cultures of endometrial stromal cells are being used to investigate the modulation of decidualization by several signaling molecules. Conventionally, human or mouse endometrial stromal cells are used. However, the availability of human samples is very often limited. Furthermore, the use of murine tissues is accompanied with variety in the method of culturing. This study presents a validated and standardized method to obtain pure Endometrial Epithelial Cell (EEC) and Stromal Cell (ESC) cultures using adult intact mice treated with estrogen for three consecutive days. The protocol is optimized to improve the yield, viability, and purity of the cells and was further extended in order to study decidualization in a coculture of EEC and ESC. This model may be suitable to exploit the importance of both cell types in decidualization and to evaluate the contribution of significant signaling molecules secreted by EEC or ESC during the intercellular communication.
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spelling pubmed-54087752017-05-12 Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization De Clercq, Katrien Hennes, Aurélie Vriens, Joris J Vis Exp Developmental Biology Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. Although many efforts have been made to reveal the underlying mechanisms of decidualization, the exact signaling between the epithelial cells that are in contact with the embryo and the underlying stromal cells remains poorly understood. Therefore, studying decidualization in a way that takes both the epithelial and stromal cells into account could improve our knowledge about the molecular details of decidualization. For this purpose, in vivo models of artificial decidualization are physiologically the most relevant; however, manipulation of intercellular communication is limited. Currently, in vitro cultures of endometrial stromal cells are being used to investigate the modulation of decidualization by several signaling molecules. Conventionally, human or mouse endometrial stromal cells are used. However, the availability of human samples is very often limited. Furthermore, the use of murine tissues is accompanied with variety in the method of culturing. This study presents a validated and standardized method to obtain pure Endometrial Epithelial Cell (EEC) and Stromal Cell (ESC) cultures using adult intact mice treated with estrogen for three consecutive days. The protocol is optimized to improve the yield, viability, and purity of the cells and was further extended in order to study decidualization in a coculture of EEC and ESC. This model may be suitable to exploit the importance of both cell types in decidualization and to evaluate the contribution of significant signaling molecules secreted by EEC or ESC during the intercellular communication. MyJove Corporation 2017-03-02 /pmc/articles/PMC5408775/ /pubmed/28287563 http://dx.doi.org/10.3791/55168 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Developmental Biology
De Clercq, Katrien
Hennes, Aurélie
Vriens, Joris
Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title_full Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title_fullStr Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title_full_unstemmed Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title_short Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
title_sort isolation of mouse endometrial epithelial and stromal cells for in vitro decidualization
topic Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408775/
https://www.ncbi.nlm.nih.gov/pubmed/28287563
http://dx.doi.org/10.3791/55168
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