Extracellular Vesicles from Follicular and Ampullary Fluid Isolated by Density Gradient Ultracentrifugation Improve Bovine Embryo Development and Quality.
Asaadi, Anise; Dolatabad, Nima Azari; Atashi, Hadiet al.
2021 • In International Journal of Molecular Sciences, 22 (2), p. 1 - 16
[en] 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.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Asaadi, Anise ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium ; Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University, Shiraz 7196484334, Iran
Dolatabad, Nima Azari ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium
Atashi, Hadi ; Université de Liège - ULiège > Département GxABT > Animal Sciences (AS) ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium ; Department of Animal Science, Shiraz University, Shiraz 7144165186, Iran
Raes, Annelies; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium
Van Damme, Petra; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium
Hoelker, Michael; Department of Animal Breeding and Husbandry, University of Bonn, 53012 Bonn, Germany
Hendrix, An ; Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, 9000 Ghent, Belgium
Pascottini, Osvaldo Bogado ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium ; Department of Veterinary Sciences, Gamete Research Center, University of Antwerp, 2610 Antwerp, Belgium
Van Soom, Ann ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium
Kafi, Mojtaba ; Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University, Shiraz 7196484334, Iran
Pavani, Krishna Chaitanya ; Department of Reproduction, Obstetrics and Herd Health, Ghent University, 9820 Merelbeke, Belgium
Language :
English
Title :
Extracellular Vesicles from Follicular and Ampullary Fluid Isolated by Density Gradient Ultracentrifugation Improve Bovine Embryo Development and Quality.
This work was supported by Ghent University (BOF GOA 2018000504 (GOA030-18 BOF)). KCP is supported by Fonds Wetenschappelijk Onderzoek: 1228821N, OBP is supported by Fonds Wetenschappelijk Onderzoek: 12Y5220N Institutional Review Board Statement: Experimental protocols performed on cows in this study We are grateful to Wim Van Den Broeck and Liesbeth Couck from the Department of Morphology-Faculty of Veterinary Medicine, Ghent University for their help with the transmission electron microscopy. The authors thank Sofie De Geyter for her excellent technical assistance.Funding: This work was supported by Ghent University (BOF GOA 2018000504 (GOA030-18 BOF)). KCP is supported by Fonds Wetenschappelijk Onderzoek: 1228821N, OBP is supported by Fonds Wetenschappelijk Onderzoek: 12Y5220N Institutional Review Board Statement: Experimental protocols performed on cows in this study were approved by the state of-fice for Nature, Environment, and Consumer Protection of North Rhine-Westphalia, Germany (Landesamt für Natur, Umwelt und Verbraucherschutz Nordrhein-Westfalen, Deutschland). The blood sample collection and ovum pick-up (OPU) procedures were approved under license numbers 84-02.04.2015.A139 and 84-02.04.2014.A500, respectively
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