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Regular Article |
a Université Catholique de Louvain, Unité des Sciences Vétérinaires
b Institut de Statistiques, B-1348 Louvain-la-Neuve, Belgium
ABSTRACT
This study compares the effects of conventional controlled-rate freezing and vitrification on the morphology and metabolism of in vitro-produced bovine blastocysts. Day 7 expanded blastocysts cultured in synthetic oviduct fluid with 5% fetal calf serum were frozen in 1.36 M glycerol, 0.25 M sucrose or vitrified in 25% glycerol, 25% ethylene glycol. Cell alterations and in vitro development were evaluated immediately after thawing or after 72 h. The effect of cryopreservation on inner cell mass and trophectoderm (TE) cell number as well as glucose, pyruvate, and oxygen uptakes, and lactate release by blastocysts were evaluated. Immediately after thawing, blastocysts showed equivalent cell membrane permeabilization after both cryopreservation procedures, while alterations in nuclear staining were more frequent in vitrified embryos. After culture, similar survival and hatching rates were observed. Both procedures decreased cell number immediately after thawing and after 72 h. However, the number of TE cells was lower in frozen embryos than in vitrified ones. In relation to this, frozen blastocysts showed a decrease in glucose, pyruvate, and oxygen uptake, although those parameters were not altered in vitrified embryos. An increased glycolytic activity was also observed in frozen embryos, indicating a stress response to this procedure.
First decision: 4 December 2000.
1 Part of this work was funded by Action de Recherche Concertée de la Direction générale de la Recherche scientifique, Communauté Française de Belgique (grant number: 96/01-196) and by European Union grant QLK3-CT-1999-00104 (Quality of Life).
2 Correspondence: I. Donnay, UCL, Unité des Sciences vétérinaires, Place croix du Sud, 3, 1348 Louvain-la-Neuve, Belgium. FAX: 32 10 47 37 17;donnay{at}vete.ucl.ac.be
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