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. 2019 Apr 2;9:5538. doi: 10.1038/s41598-019-41772-3

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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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Characterization of EVs isolated by ultracentrifugation. Transmission electron microscopy analysis revealed the presence of vesicles in FBS, UC-dFBS and SBI-dFBS. StemPRO contained scarce vesicle-like structures and abundant protein aggregates, while no vesicles could be detected in UF-dFBS (A). Western blotting (B) displayed strong expression of CD71 in FBS and SBI-dFBS, no expression in UC-dFBS and UF-dFBS, whereas low expression was detected in StemPRO. Expression of HDL was seen in all samples except UF-dFBS. Nano tracking analysis (C–F) indicated that UF-dFBS had the lowest particle count. Particle distribution (C–F). (C) D10 = 10% of particles are below the size indicated as D10, D50 = 50% of particles are below the size indicated as D50, D90 = 90% of particles are below the size indicated as D90. Median particle size in FBS, dFBS and UC-FBS are in a similar range, while UF-dFBS is considerably different. (D,E) Particle concentration and distribution. FBS and SBI-dFBS showed similar particle concentrations. (F) X-axis; particle size (nm) in two size ranges; 50–200 nm and 200 + nm.