Derivation of adult canine intestinal organoids for translational research in gastroenterology

Autor: Lawrance Chandra, Wang Yuan, Martin G. Martin, Martin E. Fernandez-Zapico, Karin Allenspach, Dawn Kingsbury, Elizabeth M. Snella, Michael J. Wannemuehler, Todd Atherly, Qun Wang, Dana C. Borcherding, Yoko M. Ambrosini, Albert E. Jergens, Melissa C. Skala, Mary K. Estes, Yijun Qi, Michael J. Kimber, Jonathan P. Mochel, Agnes Bourgois-Mochel, N. Matthew Ellinwood, David K. Meyerholz
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Pathology
medicine.medical_specialty
Physiology
Plant Science
In situ hybridization
Biology
Inflammatory bowel disease
GI diseases
General Biochemistry
Genetics and Molecular Biology
Canine
Translational Research
Biomedical
03 medical and health sciences
Dogs
0302 clinical medicine
Structural Biology
In vivo
Organoid
medicine
Animals
Dog Diseases
Organoid model
Enteroid
lcsh:QH301-705.5
Ecology
Evolution
Behavior and Systematics
030304 developmental biology
0303 health sciences
Stem Cells
Gastroenterology
Histology
Cell Biology
Translational research
medicine.disease
Cystic fibrosis transmembrane conductance regulator
Intestines
Organoids
lcsh:Biology (General)
biology.protein
Immunohistochemistry
Stem cell
General Agricultural and Biological Sciences
Intestinal stem cell
030217 neurology & neurosurgery
Research Article
Developmental Biology
Biotechnology
Zdroj: BMC Biology, Vol 17, Iss 1, Pp 1-21 (2019)
BMC Biology
ISSN: 1741-7007
DOI: 10.1186/s12915-019-0652-6
Popis: 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.
Databáze: OpenAIRE
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