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Ultrastructural changes of sheep cumulus–oocyte complexes following different methods of vitrification

Published online by Cambridge University Press:  17 February 2011

Bita Ebrahimi
Affiliation:
Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, P.O. Box: 14115–111, Tehran, Iran.
Mojtaba Rezazadeh Valojerdi*
Affiliation:
Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, P.O. Box: 14115–111, Tehran, Iran. Department of Embryology, Royan Institute for Reproductive Biomedicine Research, ACECR, Tehran, Iran.
Poopak Eftekhari-Yazdi
Affiliation:
Department of Embryology, Royan Institute for Reproductive Biomedicine Research, ACECR, Tehran, Iran.
Hossein Baharvand
Affiliation:
Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran.
*
All correspondence to: Mojtaba Rezazadeh Valojerdi. Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, P.O. Box: 14115–111, Tehran, Iran. Tel: +9821 82883897. Fax: +9821 88013030. e-mail: [email protected] or [email protected]

Summary

To determine the ultrastructural changes of sheep cumulus–oocyte complexes (COCs) following different methods of vitrification, good quality isolated COCs (GV stage) were randomly divided into the non-vitrified control, conventional straw, cryotop and solid surface vitrification groups. In both conventional and cryotop methods, vitrified COCs were respectively loaded by conventional straws and cryotops, and then plunged directly into liquid nitrogen (LN2); whereas in the solid surface group, vitrified COCs were first loaded by cryotops and then cooled before plunging into LN2. Post-warming survivability and ultrastructural changes of healthy COCs in the cryotop group especially in comparison with the conventional group revealed better viability rate and good preservation of the ooplasm organization. However in all vitrification groups except the cryotop group, mitochondria were clumped. Solely in the conventional straw group, the mitochondria showed different densities and were extremely distended. Moreover in the latter group, plenty of large irregular connected vesicles in the ooplasm were observed and in some parts their membrane ruptured. Also, in the conventional and solid surface vitrification groups, cumulus cells projections became retracted from the zona pellucida in some parts. In conclusion, the cryotop vitrification method as compared with other methods seems to have a good post-warming survivability and shows less deleterious effects on the ultrastructure of healthy vitrified–warmed sheep COCs.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2011

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