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. 2017 Jun 14;7:3538. doi: 10.1038/s41598-017-03788-5

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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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Analysis of the co-culture model. (A) Anticipated behaviour of cells within the microfluidic device where ARPE-19 detached due to the invasion of HUVEC; (B) Overlaid images of ARPE-19 and HUVEC at different time points; top row: control, middle row: low glucose, bottom row: low glucose with the addition of CoCl₂; ARPE-19 is depicted in red and HUVEC in green in all cases. (C,D) Quantification of the ARPE-19 and HUVEC growth area under different medium conditions and monoculture of ARPE in DME High glucose medium. Data were analyzed using 2-way ANOVA tests (showing a significant interaction effect, p < 0.0001 for (C) and (D)). Significant difference between the ARPE monolayer (purple line) and the co-culture groups (blue, red and green lines) is observed from t = 7 and onwards (p ≤ 0.0001). Pairwise comparisons between co-culture control and low glucose & hypoxia at t = 14 h (p = 0.023), and between all co-culture groups at t = 21 and t = 28 h (p < 0.0001) are significant in (C). In (D), pairwise comparisons for low glucose and low glucose & hypoxia groups (red and green lines) at t = 14 compared to t = 0 are significant (p ≤ 0.001). N = 7.