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. 2017 Sep 20;7:11998. doi: 10.1038/s41598-017-12049-4

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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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Co-culture of MDA-MB231 tumour cells with HUVECs cells. (a) 3D reconstruction of the 2D-3D co-culture model within the microdevice after 24 h of seeding, tumour cells being grown in the 3D hydrogel matrix and HUVECs as a monolayer on top of the hydrogel in the microwells. (b–g) Assessment of the integrity of the endothelium monolayer in the co-culture system, compared to control conditions (endothelium mono-culture). b- Actin staining of a control HUVEC endothelium (mono-culture). (c) Detail of a control HUVEC endothelium (mono-culture) stained with VE-Cadherin and NucBlue®. (d) Actin staining of a HUVEC endothelium in co-culture with MDA-MB-231 tumour cells, 48 h after cell seeding. (e) Detail of a HUVEC endothelium in co-culture with MDA-MB-231 tumour cells, 48 h after seeding, stained with VE-Cadherin and NucBlue®. (f) Assessment of the integrity of the HUVEC endothelium for the co-culture after 24 and 48 h compared to control conditions (mono-culture of a HUVEC monolayer) quantified as F-actin signal area. (n = 5, p < 0.02 as calculated with Kruskal-Wallis’ Test) (g) Assessment of the HUVEC cell circularity for control (mono-culture) and co-culture conditions after 24 and 48 h. Data was normally distributed and was evaluated by means of one-way ANOVA (n = 20). Graphs show average ± SEM and magnification is 200x for all images.