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. 2021 Apr 21;11:8666. doi: 10.1038/s41598-021-87939-9

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Isolation and characterization of EVs. (A) Schematic depicting EV isolation. Cells were brought to confluence, washed, and conditioned in serum-free (SF) media for a period of 15 days. Conditioned media (CM) was collected, purified (filtration, 0.45 µm), and concentrated using ultrafiltration by centrifugation (UFC, 10 kDa molecular weight cut-off) to isolate EVs. Figure was generated using Microsoft PowerPoint (https://www.microsoft.com/en-us/microsoft-365/powerpoint). (B) Representative nanoparticle tracking analysis traces depicting particle size and concentration. (C) Quantitative analysis of modal particle size in (B). (D) Quantitative analysis of particle concentration in (B). (E) Representative transmission electron microscopy images of EVs. (F) Quantitative analysis of EV protein concentration. Results are presented as mean ± SEM. CDC-EVs (n = 10); MSC-EVs (n = 4). Statistical significance was determined using the Mann–Whitney test, *P < 0.05.