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Effects of different cryopreservation methods on post-thaw culture conditions of in vitro produced bovine embryos

Published online by Cambridge University Press:  16 March 2011

Alessandra Corallo Nicacio ,
Renata Simões ,
Fabiola Freitas de Paula-Lopes ,
Flavia Regina Oliveira de Barros ,
Maria Angelica Peres ,
Mayra Elena Ortiz D'Avila Assumpção  and
Jose Antonio Visintin
Alessandra Corallo Nicacio
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil. College of Veterinary Medicine, State University of Maranhão, São Luis, Brazil.
Renata Simões
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Fabiola Freitas de Paula-Lopes
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Flavia Regina Oliveira de Barros
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Maria Angelica Peres
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Mayra Elena Ortiz D'Avila Assumpção
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Jose Antonio Visintin*
Affiliation:
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, Avenida Prof. Dr. Orlando Marques de Paiva, 87–Cidade Universitária–05508–270–São Paulo, SP, Brazil.
*
All correspondence to: José Antonio Visintin. Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo, Avenida Prof. Dr. Orlando Marques de Paiva, 87–Cidade Universitária–05508–270–São Paulo, SP, Brazil. Tel: +55 11 3091 7915. Fax: +55 11 3091 7412. e-mail: visintin@usp.br
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Summary

The aim of this work was to evaluate the effect of cryopreservation protocols on subsequent development of in vitro produced bovine embryos under different culture conditions. Expanded in vitro produced blastocysts (n = 600) harvested on days 7–9 were submitted to controlled freezing [slow freezing group: 10% ethylene glycol (EG) for 10 min and 1.2°C/min cryopreservation]; quick-freezing [rapid freezing group: 10% EG for 10 min, 20% EG + 20% glycerol (Gly) for 30 s]; or vitrification [vitrification group: 10% EG for 10 min, 25% EG + 25% Gly for 30 s] protocols. Control group embryos were not exposed to cryoprotectant or cryopreservation protocols and the hatching rate was evaluated on day 12 post-insemination. In order to evaluate development, frozen–thawed embryos were subjected to granulosa cell co-culture in TCM199 or SOFaa for 4 days. Data were analyzed by PROC MIXED model using SAS Systems for Windows®. Values were significant at p < 0.05. The hatching rate of the control group was 46.09%. In embryos cultured in TCM199, slow freezing and vitrification group hatching rates were 44.65 ± 5.94% and 9.43 ± 6.77%, respectively. In embryos cultured in SOFaa, slow freezing and vitrification groups showed hatching rates of 11.65 ± 3.37 and 8.67 ± 4.47%, respectively. In contrast, the rapid freezing group embryos did not hatch, regardless of culture medium. The slow freezing group showed higher hatching rates than other cryopreservation groups. Under such conditions, controlled freezing (1.2°C/min) can be an alternative to cryopreservation of in vitro produced bovine embryos.

Keywords

Bovine embryosCryopreservationEmbryo cultureIn vitro productionSlow freezing
Type
Research Article
Information
Zygote , Volume 20 , Issue 2 , May 2012 , pp. 117 - 122
Copyright
Copyright © Cambridge University Press 2011

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