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. 2021 Nov 9;10(11):1154. doi: 10.3390/biology10111154

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Characterization of FF EVs and JAr EVs. (A) Size profile and concentration of EVs measured by NTA. N = 3 of each of the sized follicles. Error bars display mean ± standard deviation (SD). (B) Western blot analysis of FF EVs for EV-specific marker. The presence of specific EV marker CD63 in EVs samples confirmed the successful isolation of EVs from FF. The apoA-I marker was used as a purity control for EVs and a strong signal of apoA-I was observed in unpurified FF samples compared to EVs, which indicates that EVs purified from FF by SEC had little or no contamination where ApoA-I indicates the purity of EVs. (C) EVs purified from JAr-conditioned medium showed a strong positive signal for EV-specific markers CD63, CD81 and CD9 compared to unpurified JAr-conditioned medium, which showed the enrichment of EVs compared to unpurified samples. (D) EVs purified from bovine FF were analyzed by TEM, where the black arrow indicates the typical cup-shaped of EVs. (E) EVs purified from JAr-conditioned medium were confirm and characterized by TEM, where the black arrow indicates the typical cup-shaped of EVs.