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. 2019 Dec 10;9:18756. doi: 10.1038/s41598-019-55128-4

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Transplanted ENCDCs survive, engraft, and migrate in the aganglionic colon. Two weeks after DT injection, tdT + neurospheres were transplanted into the colon wall and analysis was performed at 3 weeks after cell transplantation. Transplanted tdT + neurospheres (a, open arrow) survive in the aganglionic segment of the colon (a, closed arrows mark endogenous ganglia). Transplanted cells consist of Tuj1 + neurons (b-b”, open arrows) and Tuj1-negative neural crest-derived cells (b-b’, arrowheads). Transplanted tdT + cells (c, open arrows) are found near endogenous ganglia (c, closed arrows) and project fibers within the muscle layers (d, open arrow; closed arrow marks endogenous myenteric plexus. Wnt1-tdT ENCDCs express Tuj1 (b,c,d), Hu (e), nNOS (f), ChAT (g), and p75 (h). Scale bar 50 µm (a–h).