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. 2021 Aug 5;11:15889. doi: 10.1038/s41598-021-95434-4

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© The Author(s) 2021

Open Access This 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/.

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Ex vivo organotypic culture of colonic tissue. (A) Schematic representation of ex vivo gut culture methodology and the strategy used to perform ENSC transplantation. Illustration created by UCL Medical Illustration. (B) Representative image of colonic tunica muscularis after fine dissection of the mucosal layer. (C) Representative image of tissue mounting apparatus. (D) Representative image of mounted colonic tissue segment in organotypic culture. (E–G) Representative images of cultured colonic tissues at Day 7 (E), Day 14 (F), and Day 21 (G) demonstrating the absence of contamination, and lack of gross tissue damage, within long-term organotypic cultures.