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. 2017 Sep 6;7:10636. doi: 10.1038/s41598-017-09928-1

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© The Author(s) 2017

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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Microvessel formation, permeability and αSMA expression in surrounding pericytes. (A) A microfluidic chip with two round chambers for microvessel formation flanked by side channels for pericyte seeding (all chambers with cells and gel are marked in blue). (B) Representative images of a matched sample show αSMA + pericytes (green) in the microvascular chamber after one week in culture, counterstained with Hoechst (blue). Microvascular chambers are marked with a dotted line, diameter 2.4 mm. (C) 3D rendering of a αSMA + pericyte located on the abluminal surface of a CD31-stained microvessel. Scale bar: 30 μm. (D) Left: Endothelial microvascular networks continuously expressing CD31 (red) throughout the central chamber with open entrances between pillars. These networks form both when supported with normal (top) or tumor-derived (bottom) Lin-EpCAM-CD73+CD90+ cells. Center: Fluorescent 70 kDa RITC dextran (greyscale) show perfusability of microvascular networks and higher leakage of tumor PC-supported vessels compared to normal PC-vessels after 10 minutes. Scale bars: 500 μm. Right: Vascular permeability was measured with normal (n = 8) and tumor (n = 13) Lin-EpCAM-CD73+CD90+ cells (1 to 4 technical replicates per sample). Measurements from BE132-133 are marked in yellow and BE143-144 in blue. Statistical analysis in D by Student t-test, two-tailed, for comparison of paired parametric data. All tests were two-tailed. *p < 0.05 were considered significant. See related supplementary data Figure S9.