Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

Theriogenology. 2012 Aug;78(3):576-82. doi: 10.1016/j.theriogenology.2012.03.003. Epub 2012 May 11.

Abstract

The objectives were to evaluate the reexpansion blastocoele rate, post-thaw viability, and in vitro development of canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol (GLY) or 1.5 m ethylene glycol (EG). Fifty-one in vivo-produced canine blastocysts were randomly allocated in two groups: GLY (n = 26) and EG (n = 25). After thawing, embryos from M0 were immediately stained with the fluorescent probes propidium iodide and Hoechst 33 342 to evaluate cellular viability. Frozen-thawed embryos from M3 and M6 were cultured in SOFaa medium + 10% FCS at 38.5°C under an atmosphere of 5% CO(2) with maximum humidity, for 3 and 6 days, respectively, and similarly stained. The blastocoele reexpansion rate (24 h after in vitro culture) did not differ between GLY (76.5%) and EG (68.8%). Post-thaw viable cells rate were not significantly different between GLY and EG (66.5 ± 4.8 and 57.3 ± 4.8, respectively, mean ± SEM), or among M0 (62.3 ± 5.7%), M3 (56.9 ± 6.0%), and M6 (66.5 ± 6.0%). In conclusion, canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol or 1.5 m ethylene glycol, had satisfactory blastocoele reexpansion rates, similar post-thawing viability, and remained viable for up to 6 days of in vitro culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / physiology*
  • Cryopreservation / methods
  • Cryopreservation / veterinary*
  • Cryoprotective Agents
  • Dogs / embryology*
  • Embryo Culture Techniques / veterinary
  • Embryonic Development
  • Ethylene Glycol
  • Female
  • Glycerol
  • Hot Temperature*
  • Pregnancy

Substances

  • Cryoprotective Agents
  • Ethylene Glycol
  • Glycerol