Establishment of Bovine 3D Enteroid-derived 2D monolayers

Kate Sutton, Brigid Orr, Jayne Hope, Stina Rikke Jensen, Lonneke Vervelde

Research output: Contribution to journalArticlepeer-review

Abstract

Three-dimensional (3D) intestinal enteroids are powerful in vitro models for studying intestinal biology. However, due to their closed structure direct access to the apical surface is impeded, limiting high-throughput applications of exogenous compounds and pathogens. In this study, we describe a method for generating confluent 2D enteroids from single-cell suspensions of enzymatically-dissociated ileum-derived bovine 3D enteroids. Confluent monolayers were first achieved using IntestiCult media but to establish a defined, cost-effective culture media, we also developed a bovine enteroid monolayer (BEM) medium. The monolayers cultured in BEM media proliferated extensively and formed confluent cell layers on both Matrigel-coated plastic plates and transwell inserts by day 3 of culture. The 2D enteroids maintained the epithelial cell lineages found in 3D enteroids and ileum tissue. In addition, the monolayers formed a functional epithelial barrier based on the presence of the adherens and tight junction proteins, E-cadherin and ZO-1, and electrical resistance across the monolayer was measured from day 3 and maintained for up to 7 days in culture. The method described here will provide a useful model to study bovine epithelial cell biology with ease of access to the apical surface of epithelial cells and has potential to investigate host-pathogen interactions and screen bioactive compounds.
Original languageEnglish
Article number15 (2022)
Number of pages13
JournalVeterinary Research
Volume53
Issue number1
DOIs
Publication statusPublished - 2 Mar 2022

Keywords

  • Barrier Function
  • Bovine Enteroids
  • Epithelial Cells
  • 2D Enteroids
  • 3D Enteroids
  • TEER

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