title: Oocyte pre-IVM with caffeine improves bovine embryo survival after vitrification.
abstract: Cryopreservation of in vitro produced bovine embryos is associated with significantly reduced survival rates, mainly due to insufficient quality of the embryos. Caffeine supplementation during IVM has been used to delay meiotic resumption and concomitantly also increased embryo quality. Here, we investigated the influence of pre-IVM with caffeine on oocyte maturation, intraoocyte cAMP concentration, developmental competence after IVF, and blastocyst cryotolerance. Oocytes were obtained by slicing of ovaries and were submitted to either 2 hours culture before IVM with or without caffeine (0, 1, 5, 10, 20, 30 mM), or standard IVM (no pre-IVM). Oocytes were in vitro matured and fertilized and zygotes were cultured under standard in vitro conditions until Day 8. Expanded blastocysts derived from either standard control or the 10-mM caffeine treatments were submitted to vitrification. Caffeine delayed meiotic resumption after 9-hour IVM in a concentration-dependent manner. The cAMP levels were similar before and after IVM. Matured oocytes, cleavage, and blastocyst rates were reduced in the 30-mM caffeine concentration and were similar among the other treatment groups. Number and proportion of inner cell mass and trophectoderm cells in blastocysts did not differ among treatments. Forty-eight hours after thawing, hatching rates were higher in the 10-mM caffeine group (73.8%) compared with the standard control (59.7%). Reexpansion rates and total number of cells after 48 hours were similar in both treatments. The ratio of live/total cells was higher in the caffeine treatment. These results suggest that caffeine supplementation before IVM delayed meiotic resumption and improved blastocyst quality shown in higher cryotolerance.
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany; Facultad de Ciencias Agropecuarias, Universidad de Ciencias Aplicadas y Ambientales, Bogotá, Colombia.
authors.forename: Sandra Milena
authors.initials: SM
authors.lastname: Bernal-Ulloa
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lucas-Hahn
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany.
authors.forename: Doris
authors.initials: D
authors.lastname: Herrmann
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany.
authors.forename: Klaus-Gerd
authors.initials: KG
authors.lastname: Hadeler
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Aldag
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Baulain
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institute, Mariensee, Germany. Electronic address: heiner.niemann@fli.bund.de.
authors.forename: Heiner
authors.initials: H
authors.lastname: Niemann
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002110:Caffeine / Q000494:pharmacology*; D002417:Cattle; D015925:Cryopreservation / Q000662:veterinary*; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005260:Female; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary*; D009865:Oocytes / Q000187:drug effects*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D058989:Vitrification
chemical_list: D010726:Phosphodiesterase Inhibitors; D002110:Caffeine
keywords: Bovine; Caffeine; In vitro maturation; Pre-IVM; Vitrification