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Developmental rate and ultrastructure of vitrified human pronuclear oocytes after step-wise versus direct rehydration

Isachenko, Vladimir and Montag, Markus and Isachenko, Eugenia and Nawroht, Frank and Dessole, Salvatore and Ven, Hans van der (2004) Developmental rate and ultrastructure of vitrified human pronuclear oocytes after step-wise versus direct rehydration. Human Reproduction, Vol. 19 (3), p. 660-665. eISSN 1460-2350. Article.

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DOI: 10.1093/humrep/deh151

Abstract

BACKGROUND: This study compared the viability of human pronuclear oocytes subjected to vitrification followed by post-thaw step-wise removal of cryoprotectants versus direct rehydration, in terms of their subsequent in vitro survival and ultrastructural features. METHODS: A total of 115 three-pronuclei stage oocytes were cryopreserved in super-open-pulled straws by vitrification in 40% ethylene glycol + 0.75 mol/l sucrose for either 1 min or 10 s at 38°C, followed by direct plunging into liquid nitrogen. After thawing, oocytes vitrified for 1 min (group 1) or 10 s (group 2) were expelled into a graded series of sucrose solutions (1.0, 0.75, 0.5, 0.25 and 0.12 mol/l) for removal of the cryoprotectant in five 2.5 min steps. A second batch of oocytes vitrified for either 1 min (group 3) or 10 s (group 4) were directly expelled into culture medium at 38°C after thawing. Finally, the ultrastructural changes occurring in oocytes in each of the treatment groups were evaluated. RESULTS: Oocyte development (division to two-blastomere stage) rates after in vitroculture were 82, 83, 0 and 0% for groups 1, 2, 3 and 4, respectively. The harsh osmotic process involved in direct rehydration provoked ultrastructural changes, including the disruption of cytoplasmic and pronuclear membranes as well as intracellular organelles. CONCLUSION: The direct post-thaw rehydration of human pronuclear oocytes has lethal osmotic effects, such that protocols for vitrifying human pronuclear oocytes should include the step-wise removal of the cryoprotectant

Item Type:Article
ID Code:2316
Status:Published
Refereed:Yes
Uncontrolled Keywords:Damage, direct rehydration, human oocytes, ultrastructure, vitrification
Subjects:Area 06 - Scienze mediche > MED/40 Ginecologia e ostetricia
Divisions:001 Università di Sassari > 01 Dipartimenti > Neuroscienze, scienze materno infantili
Publisher:Oxford University Press
eISSN:1460-2350
Copyright Holders:© European Society of Human Reproduction and Embryology 2004
Deposited On:18 Aug 2009 10:07

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