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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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Cellular outgrowth onto the tissue surface. (A) Representative H&E-stained images showing cancerous (red arrows) and non-neoplastic (black arrows) cell outgrowth at different time points (0 h to 96 h). (B) A representative example of tissue slices with areas of cancerous and non-neoplastic cell outgrowth. Immunohistochemical staining showed that both types of outgrowth were positive for CK19 and negative for vimentin, indicating that outgrowth is of ductal-epithelial origin. Cancerous cell outgrowth (red dotted areas) was positive for P53 and negative for SMAD4, in contrast to the non-neoplastic one (black dotted areas), which was negative for P53 and positive for SMAD4. (C) Comparative analysis of cancerous, non-neoplastic, and global outgrowths at the indicated time points (duplicate slices from each culture, n = 8; Friedman test followed by Dunn’s multiple comparison, p ≤ 0.05; *indicates significant difference compared to 24 h time point). Time point 0 h was omitted from analysis due to the absence of any outgrowth.