title: Developmental kinetics of in vitro-produced bovine embryos: An aid for making decisions.
abstract: Embryo developmental kinetics and embryo survival after cryopreservation have been correlated with embryo quality and viability. The main objectives of this work were to analyze developmental ability and quality of in vitro-produced bovine embryos in relation to their kinetics and to establish a criterion of quality to predict further viability. Embryos were classified and grouped by their specific stage of development (2, 3-4, or ≥ 5 cells) at 44 hours post insemination (hpi) and cultured separately up to Day 8. On Days 7 and 8, good quality expanded blastocysts were vitrified or frozen. Cryopreserved surviving hatched embryos were stained for cell counts. Embryos at a more advanced stage (3-4 cells, and ≥5 cells) developed to morulae (P < 0.001) and blastocysts (P < 0.01) at higher rates than those embryos that had cleaved once by 44 hpi. Vitrification improved the hatching rates of blastocysts at 48 hours (P < 0.001) when compared with slow-rate freezing within each group of embryos (3-4 cells and ≥5 cells). After vitrification/warming, blastocysts coming from 3- to 4-cell embryos had higher hatching rates at 48 hours than those that came from ≥5-cell embryos. With regard to differential cell counts, no effect of the initial developmental stage was observed after warming/thawing. However, trophectoderm and total cells were higher in vitrified/warmed than in the frozen/thawed embryos (P < 0.001). These data show that selecting IVF embryos at 44 hpi, after the evaluation of their in vitro embryo development, could be used as noninvasive markers of embryo developmental competence and may help to select IVF embryos that would be more suitable for cryopreservation.
authors.affiliation: Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Asturias, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Carrocera
authors.affiliation: Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Asturias, Spain.
authors.forename: J N
authors.initials: JN
authors.lastname: Caamaño
authors.affiliation: Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Asturias, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Trigal
authors.affiliation: Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Asturias, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Martín
authors.affiliation: Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Asturias, Spain. Electronic address: mcdiez@serida.org.
authors.forename: C
authors.initials: C
authors.lastname: Díez
mesh_terms: D000818:Animals; D001755:Blastocyst; D002417:Cattle; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D003657:Decision Making; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005615:Freezing; D007700:Kinetics; D008297:Male
keywords: Bovine embryo; Cryopreservation; Embryo kinetics; Embryo quality; In vitro production