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. 2019 Oct 10;10:4602. doi: 10.1038/s41467-019-12373-5

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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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Fig. 3 — Enhancement of vascularization and anastomoses in subcutaneous space of SCID-Beige mice. a Schematics and a digital photo of a Mesh device, which is a diffusion chamber with HUVEC meshes (~25 μm thick, purple) in the fibrin gel (gray) on the top and bottom. The diffusion chamber is a cylindrical cell container with a PDMS ring (blue) as the wall and two nylon grids (green) as the top and bottom. The dimensions of each component are labeled in the schematic. b Fluorescent images of randomly mixed cells (top) and microvascular mesh (bottom) placed on diffusion chambers after 2 days of culture in EGM-2 medium. HUVEC:NHDF = 9:1, Human CD31 antibody is green to show HUVEC, α-smooth muscle actin (α-SMA) antibody is red to show NHDF, and nylon grid is blue. c Cross-sectional hematoxylin/eosin staining images of retrieved devices after 14 days of implantation. Yellow arrowheads point to blood vessels with erythrocytes inside. d Density and area percentage of blood vessels at the interface between the device and panniculus carnosus muscle. n = 6 in the No cell and Random, and n = 8 in Mesh groups. Data are mean ± SEM; **P < 0.01, ***P < 0.001, NS (P > 0.05) no significant difference. One-way analysis of variance. e Cross-sectional immunostaining images of human (red) and mouse (green) CD31 antibodies showing the human and mouse blood vessels at the interface between the device and panniculus carnosus muscle. f Confocal images of perfused lectins bound to human (UEA-I, green) and mouse (GSL-I, red) endothelial cells, confirming the anastomoses between human and mouse vessels. g Blood-perfused human vasculatures anastomosed with mouse vascular system in Mesh device after 10 days of subcutaneous implantation. HUVEC-GFP is green and perfused dye DiI in vessels is red. h Representative immunostaining images of mature human vasculatures (human CD31 antibody is green) covered with perivascular cells (α-SMA antibody is red) in retrieved Random and Mesh device after 10 days of subcutaneous implantation. i The percentage of perivascular cell (PVC) coverage is 19 ± 9% (n = 3; mean ± SEM) and 65 ± 6% (n = 6) for the Random and Mesh devices, respectively. **P < 0.01. Unpaired two-tailed t test