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ORIGINAL RESEARCH article

Front. Vet. Sci.
Sec. Comparative and Clinical Medicine
Volume 11 - 2024 | doi: 10.3389/fvets.2024.1455262

Assessment of equine intestinal epithelial junctional complexes and barrier permeability using a monolayer culture system

Provisionally accepted
Amy Stewart Amy Stewart 1Jamie Kopper Jamie Kopper 2*Caroline McKinney-Aguirre Caroline McKinney-Aguirre 1Brittany Veersammy Brittany Veersammy 1Dipak Kumar Sahoo Dipak Kumar Sahoo 2John M. Freund John M. Freund 1Liara Gonzalez Liara Gonzalez 1
  • 1 College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, United States
  • 2 College of Veterinary Medicine, Iowa State University, Ames, United States

The final, formatted version of the article will be published soon.

    Gastrointestinal disease is a leading cause of death in mature horses. A lack of in vitro modeling has impeded the development of novel therapeutics. The objectives of this study were to develop and further characterize a small intestinal monolayer cell culture derived from equine jejunum including establishing normal measurements of intestinal permeability and restitution. Three-dimensional enteroids, derived from postmortem sampling of equine jejunum, were utilized to develop confluent epithelial monolayers. The presence of differentiated intestinal epithelial cell types and tight junctions were confirmed using histology, reverse transcription PCR (RT-PCR), RNAscope, protein immunofluorescence and transmission electron microscopy. Transepithelial resistance (TER) and macromolecule flux were assessed as measurements of paracellular and transcellular permeability. Scratch assays were utilized to model and assess intestinal restitution. Monolayer cell cultures reached 100% confluency by ~ 5-7 days. Equine jejunum monolayers were confirmed as epithelial in origin, with identification of differentiated intestinal epithelial cell types and evidence of tight junction proteins. Function of the intestinal barrier was supported by acquisition of physiologically normal TER values (179.9 +/-33.7 ohms*cm 2 ) and limited macromolecule flux (22±8.8% at 60 min). Additionally, following a scratch wound, epithelial cell monolayers migrated to close gap defects within 24 hours. In conclusion, this study describes the development of a novel intestinal epithelial monolayer cell culture for equine jejunum, and provides evidence of intestinal epithelial cell differentiation, formation of physiologically relevant barrier function and use as a model of intestinal restitution to test potential therapeutics for equine colic.

    Keywords: claudin, horse, Jejunum, Occludin, Scratch assay, Transepithelial resistance

    Received: 26 Jun 2024; Accepted: 06 Sep 2024.

    Copyright: © 2024 Stewart, Kopper, McKinney-Aguirre, Veersammy, Sahoo, Freund and Gonzalez. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Jamie Kopper, College of Veterinary Medicine, Iowa State University, Ames, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.