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Effect of vitrification technique and assisted hatching on rabbit embryo developmental rate

Published online by Cambridge University Press:  01 February 2009

M. Popelková ,
Z. Turanová ,
L. Koprdová ,
A. Ostró ,
S. Toporcerová ,
A. V. Makarevich  and
P. Chrenek
M. Popelková
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
Z. Turanová
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
L. Koprdová
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic. Department of Zoology and Anthropology, Faculty of Nature Sciences, Constantine the Philosopher University, Nitra, Slovak Republic.
A. Ostró
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
S. Toporcerová
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
A. V. Makarevich
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
P. Chrenek*
Affiliation:
RIAP SARC, Hlohovská 2, 949 92 Nitra, Slovak Republic. Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic. Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
*
All correspondence to: Peter Chrenek. RIAP SARC, Hlohovská 2, 949 92 Nitra, Slovak Republic. Tel: +421 37 6546 236. Fax: +421 37 6546 189. e-mail: [email protected]
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Summary

The aim of the study was to determine the efficiency of two vitrification techniques followed by two assisted hatching (AH) techniques based on post-thaw developmental capacity of precompacted rabbit embryos and their ability to leave the zona pellucida (hatching) during in vitro culture. The total cell number and embryo diameter as additional markers of embryo quality after warming were evaluated. In vivo fertilized, in vitro cultured 8–12-cell rabbit embryos obtained from superovulated rabbit does were cryopreserved by two-step vitrification method using ethylene glycol (EG) as cryoprotectant or by one-step vitrification method with EG and Ficoll (EG+Ficoll). Thawed embryos were subjected to enzymatic or mechanical AH. Vitrified EG group showed significantly lower (P < 0.05) blastocyst rate (22.5%) and hatching rate (15%) than those vitrified with EG + Ficoll (63 and 63% resp.) and that of control (97 and 97% respectively). Significantly lower values of total cell number (P < 0.05) as well as embryo diameter (P < 0.01) in EG group compared with EG + Ficoll and control group were recorded. No significant difference was found in developmental potential of warmed embryos treated by either mechanical or enzymatic AH. The present study demonstrates that the EG + Ficoll vitrification protocol provides superior embryo survival rates over the EG vitrification protocol for 8–12-cell stage precompacted rabbit embryos. No positive effect of either mechanical or enzymatic AH on the post-thaw viability and quality of rabbit embryos in vitro was observed.

Keywords

Assisted hatchingCell numberEmbryo diameterRabbit embryoVitrification
Type
Research Article
Information
Zygote , Volume 17 , Issue 1 , February 2009 , pp. 57 - 61
Copyright
Copyright © Cambridge University Press 2008

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