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. 2019 May 24;9:7797. doi: 10.1038/s41598-019-44113-6

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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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In vitro evaluation of the therapeutic potential of pre-vascularized 3D skin substitutes. (a–e) HUVEC (0.2 × 10⁵ cells/insert) mixed with FN-G-coated NHDF were cocultured, subsequently covered with HEKn, then cultured for up to an additional 7 days. (a) Macroscopic view of the construct in the culture insert. Scale bar: 10 mm. (b–e) Histological and immunohistochemical staining with hematoxylin and eosin (b), Masson’s trichrome (c), anti-laminin 5 (d, arrows = basement membrane), and anti-CD31 (e, arrows = CD31⁺ blood vessel). Scale bars: 500 μm (b), 100 μm (c–e).