Abstract
Extracellular vesicles (EVs) have been isolated from follicular (FF) and ampullary oviduct fluid (AOF), using different isolation methods. However, it is not clear whether different purification methods can affect the functionality of resulting EVs. Here, we compared two methods (OptiPrep™ density gradient ultracentrifugation (ODG UC) and single-step size exclusion chromatography (SEC) (qEV IZON™ single column)) for the isolation of EVs from bovine FF and AOF. Additionally, we evaluated whether the addition of EVs derived either by ODG UC or SEC from FF or AOF during oocyte maturation would yield extra benefits for embryo developmental competence. The characterization of EVs isolated using ODG UC or SEC from FF and AOF did not show any differences in terms of EV sizes (40–400 nm) and concentrations (2.4 ± 0.2 × 1012−1.8 ± 0.2 × 1013 particles/mL). Blastocyst yield and quality was higher in groups supplemented with EVs isolated from FF and AOF by ODG UC, with higher total cell numbers and a lower apoptotic cell ratio compared with the other groups (p < 0.05). Supplementing in vitro maturation media with EVs derived by ODG UC from AOF was beneficial for bovine embryo development and quality.
Highlights
Over the past decades, assisted reproductive technologies (ARTs) have become an important tool to understand early embryonic development in mammals and can be applied to treat human and animal infertility and to preserve gametes from animals of high genetic merit, such as in endangered species conservation [1]
The characterization of Extracellular vesicles (EVs) isolated from ampulla oviductal fluid (AOF) and follicular fluid (FF) by both OptiPrepTM density gradient ultracentrifugation (ODG UC) and size exclusion chromatography (SEC) demonstrated equal efficacy
Our study demonstrated that the ODG UC EV isolation method yielded EVs with a higher purity, which had a significantly better functional impact on embryo development and quality compared to EVs isolated by single-step size exclusion chromatography using qEV IZONTM single column
Summary
Over the past decades, assisted reproductive technologies (ARTs) have become an important tool to understand early embryonic development in mammals and can be applied to treat human and animal infertility and to preserve gametes from animals of high genetic merit, such as in endangered species conservation [1]. Antral follicles in the mammalian ovary are composed of theca, granulosa, and cumulus cells, and the oocyte, and are filled with follicular fluid (FF) [5]. Reciprocal communication between these cells and FF is regulated by endocrine and paracrine signaling factors and results in the exchange of nutrients, antioxidants, and growth molecules, amongst others. This cooperation provides a favorable environment that is vital for the normal development of a fertile oocyte [6]. Extracellular vesicles (EVs) were identified in FF and ampulla oviductal fluid (AOF) [7,8], whereby FF-derived EVs have been described to play a crucial role in follicular development
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call