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Structural, metabolic and developmental evaluation of ovulated rabbit oocytes before and after cryopreservation by vitrification and slow freezing
Authors:Pascal Salvetti  Samuel Buff  Nathalie Daniel  Thierry Joly
Institution:a Université de Lyon, France; Unité CRYOBIO, UPSP VetAgro Sup/ISARA-Lyon, 1 avenue Bourgelat, F-69280, Marcy l'Etoile, France
b USC INRA-INSERM-UCB Lyon1, PrimaStem, INSERM U846, 18 Avenue du Doyen Lépine, F-69675 Bron, France
c INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du développement et reproduction, Jouy en Josas, F-78350, France
Abstract:The cryopreservation of oocytes is valuable for the preservation of women's fertility and might also be an interesting tool to preserve animal genetic biodiversity but it is not often used because of the very poor fertility recovered after thawing, especially in rabbit species. The objective of our study was to evaluate the effect of slow-freezing and vitrification on the structural integrity of ovulated rabbit oocytes, their ATP contents, and their developmental competence. Results show that, whatever the method is used, cryopreservation has a dramatic effect on the metabolic integrity, the structural integrity, and the developmental ability of the oocytes. Vitrification and slow freezing both impair the rabbit oocytes viability after thawing but the processes act differently. Further studies are needed to improve the cryopreservation techniques in rabbit species. Moreover, we underlined that morphology and maintenance of the structural integrity of the oocytes are not suitable enough to assess the potential for further development of cryopreserved MII oocytes. The assessment of ATP metabolism allows efficient evaluation of the viability of the frozen or vitrified oocytes. It should be used in addition to parthenogenesis to better assess the potential for further development.
Keywords:Rabbit MII oocyte  Cryopreservation  ATP assay  Confocal microscopy  Parthenogenesis
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