Despite the great scientific investments, boar semen cryopreservation still an important challenge for science, regarding post-thawed boar semen quality, regardless pre-cryopreservation quality or cryopreservation protocol used. Some proteins have been described as freezeability biological marks of boar semen. However, seminal plasma have a wide range 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 freezeability. 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 lead us to postulate whether cryopreservation with or without holding time change the interaction between seminal plasma molecules and spermatozoa, and then freezeability. Thus, three experiments will be conducted in order to understand the influence of holding time on increasing boar spermatozoa cryotolerance. First experiment aims to determine the optimal holding time to boar semen cryopreservation. Second experiment aims to study the interaction between holding time use and boar spermatozoa freezeability. Finally, the third experiment aims to elucidated, through metabolomics, which sperm or seminal molecule contributes for the increasing in boar spermatozoa freezeability.
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