Abstract
BackgroundLarge 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.ResultsOrganoids 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.ConclusionsWe 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.
Highlights
Large animal models, such as the dog, are increasingly being used for studying diseases including gastrointestinal (GI) disorders
Development of 3D cultures of canine enteroids and colonoids Using isolated canine intestine crypts, which contain adult intestinal stem cells (ISC) at the crypt base, we developed 3D intestinal enteroids and colonoids from 28 healthy and 12 diseased dogs, including dogs diagnosed with inflammatory bowel disease (IBD) (N = 9) and intestinal tumors (N = 2)
A summary of the demographics of dogs used for canine intestinal stem cell isolation, culture, and maintenance is presented in Additional file 1: Table S1
Summary
Large animal models, such as the dog, are increasingly being used for studying diseases including gastrointestinal (GI) disorders. Given the high failure rate of drugs from discovery and development through the clinical trial phase (i.e., more than 90%), there is a critical need for better animal models for preclinical studies [4]. Large animal models, such as the dog, are typically more representative than mice as they have a relatively large body size, longer life span, more closely resemble human GI physiology, and develop spontaneous, analogous diseases including inflammatory bowel disease (IBD) and colorectal cancer (CRC) [4]. The ultimate goal of our research is to culture canine intestinal organoids from diseased dogs to develop better therapeutic strategies and personalized medicine for both animal and human health
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
