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Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing |
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| Authors: | Guaitolini C Renato de Freitas Taffarel M Onghero Teixeira N Soares Sudano M José Freitas P Maria Coletto Lopes M Denise Landin-Alvarenga F da Cruz de Oliveira C Alvarenga Luz M Rezende |
| Institution: | a Laboratory of Animal Reproduction, Federal University of Espirito Santo, Brazil, UFES, Department of Veterinary Medicine, Alegre Campus, Alto Universitário s/n, Caixa Postal 16, Alegre, ES, Brazil, 29500-000 b São Paulo State University, Unesp, School of Veterinary Medicine and Animal Science, FMVZ, Department of Animal Reproduction and Veterinary Radiology Rubião Jr. s/n°, Botucatu, SP, Brazil, 18618-970 c Federal University of Minas Gerais, UFMG, Veterinary School, Department of Veterinary Clinics and Surgery, Sector of Animal Reproduction, Av Antonio Carlos, 6627, Caixa Postal 567, Pampulha Campus, Belo Horizonte, Minas Gerais, Brazil, 30123-970 d Laboratory of Hormonal Assays, Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, USP, São Paulo, SP, Brazil, 05508-270 |
| 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% 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 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. |
| Keywords: | Cryopreservation Embryo viability Blastocoele reexpansion Glycerol Ethylene glycol Dog |
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