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Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium
The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5489603/ https://www.ncbi.nlm.nih.gov/pubmed/28108942 http://dx.doi.org/10.1007/s10439-017-1797-5 |
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author | Gnecco, Juan S. Pensabene, Virginia Li, David J. Ding, Tianbing Hui, Elliot E. Bruner-Tran, Kaylon L. Osteen, Kevin G. |
author_facet | Gnecco, Juan S. Pensabene, Virginia Li, David J. Ding, Tianbing Hui, Elliot E. Bruner-Tran, Kaylon L. Osteen, Kevin G. |
author_sort | Gnecco, Juan S. |
collection | PubMed |
description | The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10439-017-1797-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5489603 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-54896032017-07-03 Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium Gnecco, Juan S. Pensabene, Virginia Li, David J. Ding, Tianbing Hui, Elliot E. Bruner-Tran, Kaylon L. Osteen, Kevin G. Ann Biomed Eng Reproductive Tissue Engineering The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10439-017-1797-5) contains supplementary material, which is available to authorized users. Springer US 2017-01-20 2017 /pmc/articles/PMC5489603/ /pubmed/28108942 http://dx.doi.org/10.1007/s10439-017-1797-5 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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. |
spellingShingle | Reproductive Tissue Engineering Gnecco, Juan S. Pensabene, Virginia Li, David J. Ding, Tianbing Hui, Elliot E. Bruner-Tran, Kaylon L. Osteen, Kevin G. Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title | Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title_full | Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title_fullStr | Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title_full_unstemmed | Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title_short | Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium |
title_sort | compartmentalized culture of perivascular stroma and endothelial cells in a microfluidic model of the human endometrium |
topic | Reproductive Tissue Engineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5489603/ https://www.ncbi.nlm.nih.gov/pubmed/28108942 http://dx.doi.org/10.1007/s10439-017-1797-5 |
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