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The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions

Gastrointestinal (GI) infections in sheep have significant implications for animal health, welfare and productivity, as well as being a source of zoonotic pathogens. Interactions between pathogens and epithelial cells at the mucosal surface play a key role in determining the outcome of GI infections...

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Autores principales: Smith, David, Price, Daniel R. G., Burrells, Alison, Faber, Marc N., Hildersley, Katie A., Chintoan-Uta, Cosmin, Chapuis, Ambre F., Stevens, Mark, Stevenson, Karen, Burgess, Stewart T. G., Innes, Elisabeth A., Nisbet, Alasdair J., McNeilly, Tom N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8456012/
https://www.ncbi.nlm.nih.gov/pubmed/34568096
http://dx.doi.org/10.3389/fcimb.2021.733811
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author Smith, David
Price, Daniel R. G.
Burrells, Alison
Faber, Marc N.
Hildersley, Katie A.
Chintoan-Uta, Cosmin
Chapuis, Ambre F.
Stevens, Mark
Stevenson, Karen
Burgess, Stewart T. G.
Innes, Elisabeth A.
Nisbet, Alasdair J.
McNeilly, Tom N.
author_facet Smith, David
Price, Daniel R. G.
Burrells, Alison
Faber, Marc N.
Hildersley, Katie A.
Chintoan-Uta, Cosmin
Chapuis, Ambre F.
Stevens, Mark
Stevenson, Karen
Burgess, Stewart T. G.
Innes, Elisabeth A.
Nisbet, Alasdair J.
McNeilly, Tom N.
author_sort Smith, David
collection PubMed
description Gastrointestinal (GI) infections in sheep have significant implications for animal health, welfare and productivity, as well as being a source of zoonotic pathogens. Interactions between pathogens and epithelial cells at the mucosal surface play a key role in determining the outcome of GI infections; however, the inaccessibility of the GI tract in vivo significantly limits the ability to study such interactions in detail. We therefore developed ovine epithelial organoids representing physiologically important gastric and intestinal sites of infection, specifically the abomasum (analogous to the stomach in monogastrics) and ileum. We show that both abomasal and ileal organoids form self-organising three-dimensional structures with a single epithelial layer and a central lumen that are stable in culture over serial passage. We performed RNA-seq analysis on abomasal and ileal tissue from multiple animals and on organoids across multiple passages and show the transcript profile of both abomasal and ileal organoids cultured under identical conditions are reflective of the tissue from which they were derived and that the transcript profile in organoids is stable over at least five serial passages. In addition, we demonstrate that the organoids can be successfully cryopreserved and resuscitated, allowing long-term storage of organoid lines, thereby reducing the number of animals required as a source of tissue. We also report the first published observations of a helminth infecting gastric and intestinal organoids by challenge with the sheep parasitic nematode Teladorsagia circumcincta, demonstrating the utility of these organoids for pathogen co-culture experiments. Finally, the polarity in the abomasal and ileal organoids can be inverted to make the apical surface directly accessible to pathogens or their products, here shown by infection of apical-out organoids with the zoonotic enteric bacterial pathogen Salmonella enterica serovar Typhimurium. In summary, we report a simple and reliable in vitro culture system for generation and maintenance of small ruminant intestinal and gastric organoids. In line with 3Rs principals, use of such organoids will reduce and replace animals in host-pathogen research.
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spelling pubmed-84560122021-09-23 The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions Smith, David Price, Daniel R. G. Burrells, Alison Faber, Marc N. Hildersley, Katie A. Chintoan-Uta, Cosmin Chapuis, Ambre F. Stevens, Mark Stevenson, Karen Burgess, Stewart T. G. Innes, Elisabeth A. Nisbet, Alasdair J. McNeilly, Tom N. Front Cell Infect Microbiol Cellular and Infection Microbiology Gastrointestinal (GI) infections in sheep have significant implications for animal health, welfare and productivity, as well as being a source of zoonotic pathogens. Interactions between pathogens and epithelial cells at the mucosal surface play a key role in determining the outcome of GI infections; however, the inaccessibility of the GI tract in vivo significantly limits the ability to study such interactions in detail. We therefore developed ovine epithelial organoids representing physiologically important gastric and intestinal sites of infection, specifically the abomasum (analogous to the stomach in monogastrics) and ileum. We show that both abomasal and ileal organoids form self-organising three-dimensional structures with a single epithelial layer and a central lumen that are stable in culture over serial passage. We performed RNA-seq analysis on abomasal and ileal tissue from multiple animals and on organoids across multiple passages and show the transcript profile of both abomasal and ileal organoids cultured under identical conditions are reflective of the tissue from which they were derived and that the transcript profile in organoids is stable over at least five serial passages. In addition, we demonstrate that the organoids can be successfully cryopreserved and resuscitated, allowing long-term storage of organoid lines, thereby reducing the number of animals required as a source of tissue. We also report the first published observations of a helminth infecting gastric and intestinal organoids by challenge with the sheep parasitic nematode Teladorsagia circumcincta, demonstrating the utility of these organoids for pathogen co-culture experiments. Finally, the polarity in the abomasal and ileal organoids can be inverted to make the apical surface directly accessible to pathogens or their products, here shown by infection of apical-out organoids with the zoonotic enteric bacterial pathogen Salmonella enterica serovar Typhimurium. In summary, we report a simple and reliable in vitro culture system for generation and maintenance of small ruminant intestinal and gastric organoids. In line with 3Rs principals, use of such organoids will reduce and replace animals in host-pathogen research. Frontiers Media S.A. 2021-09-08 /pmc/articles/PMC8456012/ /pubmed/34568096 http://dx.doi.org/10.3389/fcimb.2021.733811 Text en Copyright © 2021 Smith, Price, Burrells, Faber, Hildersley, Chintoan-Uta, Chapuis, Stevens, Stevenson, Burgess, Innes, Nisbet and McNeilly https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Smith, David
Price, Daniel R. G.
Burrells, Alison
Faber, Marc N.
Hildersley, Katie A.
Chintoan-Uta, Cosmin
Chapuis, Ambre F.
Stevens, Mark
Stevenson, Karen
Burgess, Stewart T. G.
Innes, Elisabeth A.
Nisbet, Alasdair J.
McNeilly, Tom N.
The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title_full The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title_fullStr The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title_full_unstemmed The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title_short The Development of Ovine Gastric and Intestinal Organoids for Studying Ruminant Host-Pathogen Interactions
title_sort development of ovine gastric and intestinal organoids for studying ruminant host-pathogen interactions
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8456012/
https://www.ncbi.nlm.nih.gov/pubmed/34568096
http://dx.doi.org/10.3389/fcimb.2021.733811
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