Cryopreservation and in vitro survival of bovine embryos
Keywords:Embryo, Bovine, Cryopreservation
AbstractThe aim of this study was to evaluate the dilution of cryoprotectant in 2 or steps of bovine in vitro-produced embryos after quick-freezing. A total of 1329 expanded blastocyst were kept in co-culture as control group or cryopreserved by 3 quick-freezing protocols. The embryos were exposed to 10% EG for 10 minutes then to 17%, 22% or 28% for 30 seconds. After loading, the straws were held in nitrogen vapor for 2 minutes and then plunged and stored in liquid nitrogen. After warming, cryoprotectants were diluted in two steps using 0.3 M sucrose and isotonic solution, or three steps using 0.3 M sucrose + 10% EG then 0.3 M sucrose and isotonic solution. Embryos were co-cultured on a granulosa cell monolayer and evaluated after 24 hours for re-expansion and at 24, 48, 72 and 96 hours of co-culture for hatching rates. The in vitro survival rates of embryos cryopreserved by the quick-freezing method and two-step cryoprotectant dilution were 1.94; 11.88 and 6.06%, for EG 17, EG22 and EG28 groups, respectively. At the three step dilution, the in vitro survival rates were 4.67; 9.90 and 10.78% for EG 17, EG22 and EG28 groups, respectively.
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Nicacio, A. C., Assumpção, M. E. O. D., Caetano, H. V. do A., Gerger, R. P. da C., Marques, M. G., Mello, M. R. B., Mendes, C. M., Milazzotto, M. P., Oliveira, V. P., Simões, R., Yamada, C., & Visintin, J. A. (2006). Cryopreservation and in vitro survival of bovine embryos. Brazilian Journal of Veterinary Research and Animal Science, 43(1), 51-56. https://doi.org/10.11606/issn.1678-4456.bjvras.2006.26517
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