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

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Guaitolini, C. Renato de Freitas [UNESP]
Taffarel, M. Onghero [UNESP]
Teixeira, N. Soares
Sudano, M. Jose [UNESP]
Freitas, P. Maria Coletto
Lopes, Maria Denise [UNESP]
Landim-Alvarenga, Fernanda da Cruz [UNESP]
Oliveira, C. Alvarenga de
Luz, M. Rezende

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Elsevier B.V.


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 MO 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 degrees C under an atmosphere of 5% CO2 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 MO (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. (C) 2012 Elsevier B.V. All rights reserved.



Cryopreservation, Embryo viability, Blastocoele reexpansion, Glycerol, Ethylene glycol, Dog

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Theriogenology. New York: Elsevier B.V., v. 78, n. 3, p. 576-582, 2012.