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. 2020 May 26;10:8686. doi: 10.1038/s41598-020-65628-3

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© The Author(s) 2020

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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Exosome Treatment in a Coculture System Leads to Differences in Cancer Cell Morphology and Area. Equal number of CAFs and OVCAR-3 were seeded on surfaces and were treated with or without exosomes. Images of coculture model were taken every 30 minutes over a 16-hour period. (A) OVCAR-3 area average and distribution were measured at initial (I) and final (F) time points using ImageJ (N = 3). (ii) CI-exosome treatment significantly increased OVCAR-3 spread area compared to (i) untreated spread areas at respective I and F time points. (iii) SEC-exosome treatment did not show any statistically significant change in spread area. (B) Average shape factor results show no statistical difference between (ii-iii) exosome treatment and (i) untreated groups at either initial (I) or final (F) time points. Interestingly, histogram distributions showed that (iii) SEC-exosomes led to a small percentage of elongated outlier OVCAR-3 cells at final time points (black) compared to initial points (gray). The highlighted red-gated region in (i-iii) represents OVCAR-3 that had shape factors less than 0.5. Specifically, we quantified that SEC-exosomes induced elongation in approximately 10% of OVCAR-3. Untreated and CI-exosomes induced elongation in approximately 3% of OVCAR-3. Statistics calculated using Student’s t-test and reported as values +/− SEM. *p < 0.05, ^##p < 0.005.