Journal Article

Derivation of adult canine intestinal organoids for translational research in gastroenterology

Qi, Yijun, Kimber, Michael, Martin, Martin, Bourgois-Mochel, Agnes, Meyerholz, David, Estes, Mary, Kingsbury, Dawn, Ambrosini, Yoko M., Skala, Melissa, Allenspach, Karin, Atherly, Todd, Yuan, Wang, Wannemuehler, Michael, Jergens, Albert E., Wang, Qun, Mochel, Jonathan P., Chandra, Lawrance, Fernandez-Zapico, Martin E., Borcherding, Dana C., Snella, Elizabeth, Ellinwood, N. Matthew

2019

Overview

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.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Animal models / Animal research models / Animals / Assaying / Biocompatibility / Biological research / Biology / Biomedical and Life Sciences / Biomedical research / Biopsy / Cancer / Canine / Carcinoma / Cell culture / Conductance / Cryopreservation / Cystic fibrosis / Disease / Dog Diseases - physiopathology / Dogs / Electron microscopy / Engineering Biology / Enteroid / Fibrosis / Gastroenterology / Gastrointestinal diseases / Genetic research / GI diseases / Histology / Host-pathogen interactions / Hybridization / Immunohistochemistry / In vivo methods and tests / Inflammatory bowel disease / Inflammatory bowel diseases / Intestinal stem cell / Intestine / Intestines / Intestines - physiology / Intestines - physiopathology / Kinases / Life Sciences / Medical research / Methods / Microscopy / Organoid model / Organoids / Organoids - physiology / Organoids - physiopathology / Physiological aspects / Physiology / Precision medicine / R&D / Research & development / Research Article / Ribonucleic acid / RNA / Rodents / Stem cell research / Stem cell transplantation / Stem cells / Stem Cells - cytology / Studies / Technology / Three dimensional models / Tissues / Toxicology / Translation / Translational research / Translational Research, Biomedical / Transmission electron microscopy / Tumors