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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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Integration of ENSC-derived donor cells across the gut wall. (A, B) Representative confocal z-stack image of donor ENSC-derived GFP⁺ (green), and DAPI (blue), within the gut wall 21-days after ex vivo transplantation. (C) Summary data showing analysis of donor cell body (GFP⁺/DAPI⁺) integration, measured as z-axis depth, across the gut wall at day-7 (D7), D14, and D21 after ex vivo transplantation (n = 4 for each timepoint). (D) Representative depth-coded image of donor cell integration at D21. Note the protrusion of donor ENSC-derived cells into the gut wall (dashed line) from the site of transplantation on the serosal surface (magenta arrowhead). Donor GFP⁺ cells and fibres were observed to have integrated longitudinally along the gut, both on the serosal surface (arrowheads) and within the tunica muscularis (arrows). Scale bars represent 50 μm (A, B), 30 μm (D). Error bars represent mean ± s.e.m. *P ≤ 0.05, by Welch’s t-test.