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. 2022 Mar 2;53:15. doi: 10.1186/s13567-022-01033-0

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Bovine 2D enteroid form tight junctions and a functional epithelial barrier. A Representative confocal images of the adherens junction protein, E-cadherin (green) and B 3X3 tile image demonstrating uniformed expression of the tight junction protein ZO-1 (red) in 2D enteroids seeded at 2.5 × 10⁴ cells/well on Matrigel coated transwell inserts in BEM/1% FBS media. C Z-stack modeling indicate the expression of E-cadherin (green) between the cells and D strong apical expression of F-actin (red) in the 2D enteroids. E EdU (magenta) uptake by E-cadherin⁺ (green) proliferating cells in 2D enteroids. Nuclei are visualised by Hoechst staining (blue). Cells were imaged on Matrigel coated transwell inserts. F Short-term, high TEER can be achieved by seeding 2.5 × 10⁵ cells/well and G a lower more stable TEER can be reached using 2.5 × 10⁴ cells/well. Data is the median of 5 (F) or 12 (E) independent experiments from 5 to 6 calves. Error bars ± 95% Confidence Interval. H Representative brightfield image of the destablistion of the 2D enteroids leading to a drop in TEER on day 12 of culture on transwell inserts.