A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine
Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessi...
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Frontiers Media S.A.
2020-06-01
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Series: | Frontiers in Cellular and Infection Microbiology |
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Online Access: | https://www.frontiersin.org/article/10.3389/fcimb.2020.00272/full |
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doaj-3257340dbe7b42cfac508c6b045bc74b |
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Article |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Thomas Roodsant Thomas Roodsant Marit Navis Ikrame Aknouch Ikrame Aknouch Ingrid B. Renes Ingrid B. Renes Ruurd M. van Elburg Dasja Pajkrt Katja C. Wolthers Constance Schultsz Constance Schultsz Kees C. H. van der Ark Kees C. H. van der Ark Adithya Sridhar Vanesa Muncan |
spellingShingle |
Thomas Roodsant Thomas Roodsant Marit Navis Ikrame Aknouch Ikrame Aknouch Ingrid B. Renes Ingrid B. Renes Ruurd M. van Elburg Dasja Pajkrt Katja C. Wolthers Constance Schultsz Constance Schultsz Kees C. H. van der Ark Kees C. H. van der Ark Adithya Sridhar Vanesa Muncan A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine Frontiers in Cellular and Infection Microbiology human organoids small intestine enteric pathogens polarized epithelium host-pathogen interactions gut barrier |
author_facet |
Thomas Roodsant Thomas Roodsant Marit Navis Ikrame Aknouch Ikrame Aknouch Ingrid B. Renes Ingrid B. Renes Ruurd M. van Elburg Dasja Pajkrt Katja C. Wolthers Constance Schultsz Constance Schultsz Kees C. H. van der Ark Kees C. H. van der Ark Adithya Sridhar Vanesa Muncan |
author_sort |
Thomas Roodsant |
title |
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine |
title_short |
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine |
title_full |
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine |
title_fullStr |
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine |
title_full_unstemmed |
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine |
title_sort |
human 2d primary organoid-derived epithelial monolayer model to study host-pathogen interaction in the small intestine |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular and Infection Microbiology |
issn |
2235-2988 |
publishDate |
2020-06-01 |
description |
Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research. |
topic |
human organoids small intestine enteric pathogens polarized epithelium host-pathogen interactions gut barrier |
url |
https://www.frontiersin.org/article/10.3389/fcimb.2020.00272/full |
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doaj-3257340dbe7b42cfac508c6b045bc74b2020-11-25T03:27:49ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882020-06-011010.3389/fcimb.2020.00272524689A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small IntestineThomas Roodsant0Thomas Roodsant1Marit Navis2Ikrame Aknouch3Ikrame Aknouch4Ingrid B. Renes5Ingrid B. Renes6Ruurd M. van Elburg7Dasja Pajkrt8Katja C. Wolthers9Constance Schultsz10Constance Schultsz11Kees C. H. van der Ark12Kees C. H. van der Ark13Adithya Sridhar14Vanesa Muncan15Department of Global Health-Amsterdam Institute for Global Health and Development, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsTytgat Institute for Intestinal and Liver Research, Amsterdam Gastroenterology Endocrinology and Metabolism, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsViroclinics Xplore, Schaijk, NetherlandsDanone Nutricia Research, Utrecht, NetherlandsDepartment of Pediatrics, Amsterdam University Medical Center (UMC), Emma Children's Hospital, University of Amsterdam, Amsterdam, NetherlandsDepartment of Pediatrics, Amsterdam University Medical Center (UMC), Emma Children's Hospital, University of Amsterdam, Amsterdam, NetherlandsDepartment of Pediatric Infectious Diseases, Amsterdam University Medical Center (UMC), Emma Children's Hospital, University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Global Health-Amsterdam Institute for Global Health and Development, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Global Health-Amsterdam Institute for Global Health and Development, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsDepartment of Medical Microbiology, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsTytgat Institute for Intestinal and Liver Research, Amsterdam Gastroenterology Endocrinology and Metabolism, Amsterdam University Medical Center (UMC), University of Amsterdam, Amsterdam, NetherlandsGut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.https://www.frontiersin.org/article/10.3389/fcimb.2020.00272/fullhuman organoidssmall intestineenteric pathogenspolarized epitheliumhost-pathogen interactionsgut barrier |