title: Reduced competence of immature and mature oocytes vitrified by Cryotop method: assessment by in vitro fertilization and parthenogenetic activation in a bovine model.
abstract: This study aimed to evaluate the embryo development competence, the nuclear maturation and the viability of germinal vesicle (GV) and metaphase II (MII) oocytes vitrified by the Cryotop method. Cumulus-oocyte complexes were derived from bovine ovaries and three experiments were conducted. In Experiment 1, GV oocytes were vitrified and underwent in vitro maturation (IVM) or not and their nuclear maturation was assessed by orcein staining. In Experiment 2, GV oocytes and MII oocytes were vitrified or not and the viability was assessed by calcein/ethidium homodimer-1 staining. In Experiment 3, MII oocytes matured before or after vitrification were submitted to in vitro fertilization (IVF) and parthenogenetic activation (PA) in order to evaluate embryo development. No difference was found for the nuclear maturation rate in the GV group (50%) and the GV control group (67%; P = 0.23) and for viability rate (56%; 77%; P = 0.055, respectively). However, in the MII group (27%) viability was significantly lower than that of the MII control group (84%; P < 0.0001). The cleavage rate by IVF and PA was similar in the GV group and the MII group. In contrast, vitrified MII oocytes showed no capacity for blastocyst development after IVF or PA and vitrified GV oocytes were able to develop to blastocysts only after PA, but not after IVF. In conclusion, oocyte vitrification by the Cryotop method reduced the capacity for embryo development. Vitrification of GV oocytes, however, did not influence the capacity of meiotic nuclear maturation and they exhibited higher viability following vitrification at the MII stage.
authors.affiliation: Department of Obstetrics and Gynecology,School of Medicine of Ribeirão Preto,University of São Paulo,Ribeirão Preto,SP,Brazil.
authors.forename: Daiane L
authors.initials: DL
authors.lastname: Bulgarelli
authors.affiliation: Department of Obstetrics and Gynecology,School of Medicine of Ribeirão Preto,University of São Paulo,Ribeirão Preto,SP,Brazil.
authors.forename: Alessandra A
authors.initials: AA
authors.lastname: Vireque
authors.affiliation: Department of Obstetrics and Gynecology,School of Medicine of Ribeirão Preto,University of São Paulo,Ribeirão Preto,SP,Brazil.
authors.forename: Caroline P
authors.initials: CP
authors.lastname: Pitangui-Molina
authors.affiliation: Department of Obstetrics and Gynecology,School of Medicine of Ribeirão Preto,University of São Paulo,Ribeirão Preto,SP,Brazil.
authors.forename: Marcos F
authors.initials: MF
authors.lastname: Silva-de-Sá
authors.affiliation: Department of Obstetrics and Gynecology,School of Medicine of Ribeirão Preto,University of São Paulo,Ribeirão Preto,SP,Brazil.
authors.forename: Ana Carolina J
authors.initials: ACJ
authors.lastname: de Sá Rosa-E-Silva
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002417:Cattle; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000187:drug effects*; D009866:Oogenesis / Q000187:drug effects / Q000502:physiology; D010312:Parthenogenesis; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents
keywords: Bovine oocyte; Embryo Development; Viability; Vitrification