Abstract
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.
| Original language | English (US) |
|---|---|
| Article number | 33 |
| Journal | BMC Biology |
| Volume | 17 |
| Issue number | 1 |
| DOIs | |
| State | Published - Apr 11 2019 |
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Keywords
- Canine
- Enteroid
- GI diseases
- Intestinal stem cell
- Organoid model
- Translational research
ASJC Scopus subject areas
- Biotechnology
- Structural Biology
- Ecology, Evolution, Behavior and Systematics
- Physiology
- Biochemistry, Genetics and Molecular Biology(all)
- Agricultural and Biological Sciences(all)
- Plant Science
- Developmental Biology
- Cell Biology
Cite this
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.
In: BMC Biology, Vol. 17, No. 1, 33, 11.04.2019.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Derivation of adult canine intestinal organoids for translational research in gastroenterology
AU - Chandra, Lawrance
AU - Borcherding, Dana C.
AU - Kingsbury, Dawn
AU - Atherly, Todd
AU - Ambrosini, Yoko M.
AU - Bourgois-Mochel, Agnes
AU - Yuan, Wang
AU - Kimber, Michael
AU - Qi, Yijun
AU - Wang, Qun
AU - Wannemuehler, Michael
AU - Ellinwood, N. Matthew
AU - Snella, Elizabeth
AU - Martin, Martin
AU - Skala, Melissa
AU - Meyerholz, David
AU - Estes, Mary
AU - Fernandez-Zapico, Martin E
AU - Jergens, Albert E.
AU - Mochel, Jonathan P.
AU - Allenspach, Karin
PY - 2019/4/11
Y1 - 2019/4/11
N2 - 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.
AB - 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.
KW - Canine
KW - Enteroid
KW - GI diseases
KW - Intestinal stem cell
KW - Organoid model
KW - Translational research
UR - http://www.scopus.com/inward/record.url?scp=85064207587&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85064207587&partnerID=8YFLogxK
U2 - 10.1186/s12915-019-0652-6
DO - 10.1186/s12915-019-0652-6
M3 - Article
C2 - 30975131
AN - SCOPUS:85064207587
VL - 17
JO - BMC Biology
JF - BMC Biology
SN - 1741-7007
IS - 1
M1 - 33
ER -