Journal
REPRODUCTION FERTILITY AND DEVELOPMENT
Volume 32, Issue 4, Pages 409-418Publisher
CSIRO PUBLISHING
DOI: 10.1071/RD19058
Keywords
fallopian tube; oocyte; spermatozoa; zona pellucida
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Funding
- Ciencias sem Fronteiras (Brazil) program [205593/2014-3]
- EU-FECUND program [312097]
- Credit incitatifs INRA CI-PHASE program
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High polyspermy is one of the major limitations of porcine in vitro fertilisation (IVF). The addition of oviductal fluid (OF) during IVF reduces polyspermy without decreasing the fertilisation rate. Because extracellular vesicles (EVs) have been described as important OF components, the aim of this study was to evaluate the effect of porcine oviductal EVs (poEVs) on IVF efficiency compared with porcine OF (fresh and lyophilised). OF was collected from abattoir oviducts by phosphate-buffered saline flush, and poEVs were isolated by serial ultracentrifugation. Four IVF treatments were conducted: poEVs (0.2 mg mL(-1)), OF (10%), lyophilized and reconstituted pure OF (LOF; 1%) and IVF without supplementation (control). Penetration, monospermy and IVF efficiency were evaluated. Transmission electron microscopy showed an EVs population primarily composed of exosomes (83%; 30-150 nm). Supplementation with poEVs during IVF increased monospermy compared with control (44% vs 17%) while maintaining an acceptable penetration rate (61% vs 78% respectively) in a similar way to OF and LOF. Western blotting revealed poEVs proteins involved in early reproductive events, including zona pellucida hardening. In conclusion, our finding show that poEVs are key components of porcine OF and may play roles in porcine fertilisation and polyspermy regulation, suggesting that supplementation with poEVs is a reliable strategy to decrease porcine polyspermy and improve in vitro embryo production outcomes.
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