Despite great scientific investments, boar semen cryopreservation still presents an important challenge for science, regarding post-thawed boar semen quality, independently of pre-cryopreservation quality or cryopreservation protocol used. Some proteins have been described as freezability biological marks of boar semen. However, seminal plasma has a wide range of components, so it lead us to believe that not only proteins but also lipids, ions, and others components could be beneficial or detrimental to boar semen freezability. Jointly, holding time (incubation time in own seminal plasma before cryopreservation) has been used to improve post-thawed boar semen quality. Holding time allows greater contact of spermatozoa with seminal plasma components, and it is reported to improve cryotolerance at low temperatures. It led us to postulate whether cryopreservation with or without holding time changes the interaction between seminal plasma molecules and spermatozoa, and then freezability. Thus, three experiments will be conducted in order to understand the influence of the holding time on increasing boar spermatozoa cryotolerance. First experiment aims to determine theoptimal holding time to boar semen cryopreservation. Second experiment aims to study the relation between boar spermatozoa freezability and holding time. Finally, the third experiment aims to elucidate, through metabolomics which seminal molecules contributefor the increasing in boar spermatozoa freezability.
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