Effect of Cryoprotectant Composition on In Vitro Viability of In Vitro Fertilized and Cloned Bovine Embryos Following Vitrification and In-Straw Dilution

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  • TANIGUCHI Masayasu
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • IKEDA Akiko
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • ARIKAWA Eri
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • WONGSRIKEAO Pimprapar
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • AGUNG Budiyanto
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • NAOI Hideaki
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University
  • NAGAI Takashi
    Department of Research Planning and Coordination, National Institute of Livestock and Grassland Science
  • OTOI Takeshige
    Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University

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In this study the efficacy of the combination of glycerol (GLY) and ethylene glycol (EG) as cryoprotectants in a vitrification method developed for direct embryo transfer was evaluated by in vitro development of in vitro fertilized (IVF) and somatic cell nuclear transfer (SCNT) embryos after vitrification. The IVF and SCNT blastocysts were vitrified in either 40% GLY, 30% GLY + 10% EG, or 20% GLY + 20% EG using French straws. After warming, the straws were held vertically for 1 min without shaking and were then placed horizontally for 5 min to dilute the cryoprotectants. After washing, the vitrified-warmed embryos were cultured in vitro for 72 h. There were no differences among the vitrification solutions with respect to the rates of vitrified-warmed IVF and SCNT embryos surviving and developing to the hatched blastocyst stage. However, the rates of development to the hatched blastocyst stage of the SCNT embryos vitrified with 40% GLY tended to be higher than those vitrified with 30% GLY + 10% EG or 20% GLY + 20% EG (26% vs. 7-8%, respectively). The development rates to the hatched blastocyst stage of the IVF and SCNT embryos vitrified with solution containing EG were significantly lower (P<0.05) than those of non-vitrified embryos. These results suggest that use of the combination of GLY and EG as cryoprotectants had no beneficial effect on the viability of embryos after in-straw dilution. However, this method is so simple that it can be used for practical direct transfer of vitrified embryos in the field.<br>

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