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. 2019 Feb 14;9:2133. doi: 10.1038/s41598-019-38603-w

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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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Tissue viability and structural integrity. (A) Representative overview images of H&E-stained whole tissue slices. See Figure S1 for high magnification images. (B) Representative H&E-stained tissue slices showing viable tissue and foci of apoptosis and necrosis. (C) Left panel, Digital annotation of non-viable tissue areas within the slice. Red annotated areas indicate non-viable tissue. Right panel, Quantification of non-viable tissue areas during culture time (n = 12; Friedman test followed by Dunn’s multiple comparison; p > 0.05). In the violin plot, the central band, box and whiskers represent median value, interquartile range, and maximum and minimum values, respectively. (D) Schematic diagram showing the annotation of a whole slice into 3 separate zones - peripheral, intermediate and central, each with a width of one-third of the tissue slice diameter. The heatmap shows spatial distribution of mean non-viable tissue areas (%) in the 3 zones with respect to culture time. The scale indicates mean values. (E) Representative transmission electron photomicrographs (n = 3) of control (0 h) and cultured slices (24 h and 72 h) depicting morphological alterations of epithelial outgrowth, cancer cells, blood vessels and stromal components during culture.