Extracellular Vesicles from Follicular and Ampullary Fluid Isolated by Density Gradient Ultracentrifugation Improve Bovine Embryo Development and Quality

• Published in: International journal of molecular sciences 2021-01, Vol.22 (2), p.578
• Main Authors: Asaadi, Anise, Dolatabad, Nima Azari, Atashi, Hadi, Raes, Annelies, Van Damme, Petra, Hoelker, Michael, Hendrix, An, Pascottini, Osvaldo Bogado, Van Soom, Ann, Kafi, Mojtaba, Pavani, Krishna Chaitanya
• Format: Article
• Language: English
• Subjects: ampullary oviductal fluid; Animals; Apoptosis; Blastocyst; Blastocyst - drug effects; Catalysis; Cattle; Centrifugation, Density Gradient; Chromatography; Computer Science Applications; Culture Media, Conditioned; Embryo Culture Techniques; Embryonic Development; Embryonic Development - genetics; Embryos; Extracellular vesicles; Extracellular Vesicles - metabolism; Female; Fluids; follicular fluid; Follicular Fluid - chemistry; Follicular Fluid - metabolism; Gametocytes; General Medicine; Hair Cells, Ampulla; Hair Cells, Ampulla - chemistry; Hair Cells, Ampulla - metabolism; Humans; Inorganic Chemistry; Life sciences; Mammals; Maturation; MicroRNAs; Molecular Biology; Médecine vétérinaire & santé animale; Oogenesis; Oogenesis - drug effects; Organic Chemistry; Oviduct; Oviducts; Oviducts - drug effects; Physical and Theoretical Chemistry; Physiology; Proteins; Sciences du vivant; Size exclusion chromatography; Ultracentrifugation; Vesicles; Veterinary medicine & animal health
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22020578

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 × 10 -1.8 ± 0.2 × 10 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 ( < 0.05). Supplementing in vitro maturation media with EVs derived by ODG UC from AOF was beneficial for bovine embryo development and quality.