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Hum. Reprod. Advance Access published online on June 2, 2005
Human Reproduction, doi:10.1093/humrep/deh805
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org
Received October 19, 2004
Revised December 14, 2004
Accepted January 19, 2005

Article

Cryoloop vitrification of rabbit oocytes

X.Y. Cai 1, G.A. Chen 1*, Y. Lian 1, X.Y. Zheng 1, and H.M. Peng 1

1 Department of Obstetrics and Gynaecology, Third Hospital, Peking University, Peking, China 100083

* To whom correspondence should be addressed.
G.A. Chen, E-mail: Chenguian{at}bjmu.edu.cn


  Abstract

BACKGROUND: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimization. In this study, rabbit oocytes (fertilized by ICSI) were vitrified with cryoloops, and the effect of three different cryopreservation protocols on spindle configuration and embryo quality was assessed. METHODS: Metaphase II rabbit oocytes were randomly assigned to one of four groups: (i) control; (ii) E40 [40% ethylene glycol (EG)]; (iii) ED20 [20% EG + 20% dimethylsulphoxide (DMSO)]; and (iv) ED20 + M (20% EG+20% DMSO+ vitrification machine). After warming, one part of each group was fertilized by ICSI to examine the fertilization and embryo cleavage ability, and the others were immunostained for tubulin and chromatin before visualization using confocal microscopy. RESULTS: The survival rates after warming were 79.1, 83.1 and 82.3%, respectively. In protocols E40 and ED20, the spindles were severely injured and the embryo quality not good compared with those in the ED20 + M group. CONCLUSIONS: The fastest cooling rate in combination with EG and DMSO as cryoprotectants had the fewest adverse effects on the spindle configuration of rabbit oocytes and embryo development.

Keywords: cryoloop; cryopreservation; oocyte/rabbit; spindle; vitrification.
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