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Derivation of adult canine intestinal organoids for translational research in gastroenterology
BACKGROUND: Large animal models, such as the dog, are increasingly being used for studying diseases including gastrointestinal (GI) disorders. Dogs share similar environmental, genomic, anatomical, and intestinal physiologic features with humans. To bridge the gap between commonly used animal models...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6460554/ https://www.ncbi.nlm.nih.gov/pubmed/30975131 http://dx.doi.org/10.1186/s12915-019-0652-6 |
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author | Chandra, Lawrance Borcherding, Dana C. Kingsbury, Dawn Atherly, Todd Ambrosini, Yoko M. Bourgois-Mochel, Agnes Yuan, Wang Kimber, Michael Qi, Yijun Wang, Qun Wannemuehler, Michael Ellinwood, N. Matthew Snella, Elizabeth Martin, Martin Skala, Melissa Meyerholz, David Estes, Mary Fernandez-Zapico, Martin E. Jergens, Albert E. Mochel, Jonathan P. Allenspach, Karin |
author_facet | Chandra, Lawrance Borcherding, Dana C. Kingsbury, Dawn Atherly, Todd Ambrosini, Yoko M. Bourgois-Mochel, Agnes Yuan, Wang Kimber, Michael Qi, Yijun Wang, Qun Wannemuehler, Michael Ellinwood, N. Matthew Snella, Elizabeth Martin, Martin Skala, Melissa Meyerholz, David Estes, Mary Fernandez-Zapico, Martin E. Jergens, Albert E. Mochel, Jonathan P. Allenspach, Karin |
author_sort | Chandra, Lawrance |
collection | PubMed |
description | BACKGROUND: Large animal models, such as the dog, are increasingly being used for studying diseases including gastrointestinal (GI) disorders. Dogs share similar environmental, genomic, anatomical, and intestinal physiologic features with humans. To bridge the gap between commonly used animal models, such as rodents, and humans, and expand the translational potential of the dog model, we developed a three-dimensional (3D) canine GI organoid (enteroid and colonoid) system. Organoids have recently gained interest in translational research as this model system better recapitulates the physiological and molecular features of the tissue environment in comparison with two-dimensional cultures. RESULTS: Organoids were derived from tissue of more than 40 healthy dogs and dogs with GI conditions, including inflammatory bowel disease (IBD) and intestinal carcinomas. Adult intestinal stem cells (ISC) were isolated from whole jejunal tissue as well as endoscopically obtained duodenal, ileal, and colonic biopsy samples using an optimized culture protocol. Intestinal organoids were comprehensively characterized using histology, immunohistochemistry, RNA in situ hybridization, and transmission electron microscopy, to determine the extent to which they recapitulated the in vivo tissue characteristics. Physiological relevance of the enteroid system was defined using functional assays such as optical metabolic imaging (OMI), the cystic fibrosis transmembrane conductance regulator (CFTR) function assay, and Exosome-Like Vesicles (EV) uptake assay, as a basis for wider applications of this technology in basic, preclinical and translational GI research. We have furthermore created a collection of cryopreserved organoids to facilitate future research. CONCLUSIONS: We establish the canine GI organoid systems as a model to study naturally occurring intestinal diseases in dogs and humans, and that can be used for toxicology studies, for analysis of host-pathogen interactions, and for other translational applications. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12915-019-0652-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6460554 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-64605542019-04-22 Derivation of adult canine intestinal organoids for translational research in gastroenterology Chandra, Lawrance Borcherding, Dana C. Kingsbury, Dawn Atherly, Todd Ambrosini, Yoko M. Bourgois-Mochel, Agnes Yuan, Wang Kimber, Michael Qi, Yijun Wang, Qun Wannemuehler, Michael Ellinwood, N. Matthew Snella, Elizabeth Martin, Martin Skala, Melissa Meyerholz, David Estes, Mary Fernandez-Zapico, Martin E. Jergens, Albert E. Mochel, Jonathan P. Allenspach, Karin BMC Biol Research Article BACKGROUND: Large animal models, such as the dog, are increasingly being used for studying diseases including gastrointestinal (GI) disorders. Dogs share similar environmental, genomic, anatomical, and intestinal physiologic features with humans. To bridge the gap between commonly used animal models, such as rodents, and humans, and expand the translational potential of the dog model, we developed a three-dimensional (3D) canine GI organoid (enteroid and colonoid) system. Organoids have recently gained interest in translational research as this model system better recapitulates the physiological and molecular features of the tissue environment in comparison with two-dimensional cultures. RESULTS: Organoids were derived from tissue of more than 40 healthy dogs and dogs with GI conditions, including inflammatory bowel disease (IBD) and intestinal carcinomas. Adult intestinal stem cells (ISC) were isolated from whole jejunal tissue as well as endoscopically obtained duodenal, ileal, and colonic biopsy samples using an optimized culture protocol. Intestinal organoids were comprehensively characterized using histology, immunohistochemistry, RNA in situ hybridization, and transmission electron microscopy, to determine the extent to which they recapitulated the in vivo tissue characteristics. Physiological relevance of the enteroid system was defined using functional assays such as optical metabolic imaging (OMI), the cystic fibrosis transmembrane conductance regulator (CFTR) function assay, and Exosome-Like Vesicles (EV) uptake assay, as a basis for wider applications of this technology in basic, preclinical and translational GI research. We have furthermore created a collection of cryopreserved organoids to facilitate future research. CONCLUSIONS: We establish the canine GI organoid systems as a model to study naturally occurring intestinal diseases in dogs and humans, and that can be used for toxicology studies, for analysis of host-pathogen interactions, and for other translational applications. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12915-019-0652-6) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-11 /pmc/articles/PMC6460554/ /pubmed/30975131 http://dx.doi.org/10.1186/s12915-019-0652-6 Text en © The Author(s). 2019 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. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Chandra, Lawrance Borcherding, Dana C. Kingsbury, Dawn Atherly, Todd Ambrosini, Yoko M. Bourgois-Mochel, Agnes Yuan, Wang Kimber, Michael Qi, Yijun Wang, Qun Wannemuehler, Michael Ellinwood, N. Matthew Snella, Elizabeth Martin, Martin Skala, Melissa Meyerholz, David Estes, Mary Fernandez-Zapico, Martin E. Jergens, Albert E. Mochel, Jonathan P. Allenspach, Karin Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title | Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title_full | Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title_fullStr | Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title_full_unstemmed | Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title_short | Derivation of adult canine intestinal organoids for translational research in gastroenterology |
title_sort | derivation of adult canine intestinal organoids for translational research in gastroenterology |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6460554/ https://www.ncbi.nlm.nih.gov/pubmed/30975131 http://dx.doi.org/10.1186/s12915-019-0652-6 |
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