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Hum. Reprod. Advance Access published online on October 20, 2005

Human Reproduction, doi:10.1093/humrep/dei346
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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@oxfordjournals.org
Received June 7, 2005
Revised August 23, 2005
Accepted September 1, 2005

Article

Clinical outcome of oocyte cryopreservation after slow cooling with a protocol utilizing a high sucrose concentration

A. Borini 1*, R. Sciajno 1, V. Bianchi 1, E. Sereni 1, C. Flamigni 2, and G. Coticchio 1

1 Tecnobios Procreazione, Via Dante 15, Italy
2 University of Bologna, 40125 Bologna, Italy

* To whom correspondence should be addressed.
A. Borini, E-mail: borini{at}tecnobiosprocreazione.it


   Abstract

BACKGROUND: Recently, interest in oocyte cryopreservation has steadily increased. Newly developed protocols have dramatically improved survival rates, removing perhaps the major hurdle that has prevented this approach from becoming a fully established form of treatment. However, the clinical efficiency of these protocols has not been exhaustively explored and therefore remains controversial. METHODS: Morphologically normal oocytes displaying the first polar body were frozen-thawed with a slow cooling protocol that utilized 1.5mol/l propane-1,2-diol (PrOH) and 0.3mol/l sucrose. RESULTS: A total of 927 oocytes from 146 patients were frozen-thawed, achieving a 74.1% survival rate. Over 76% of microinjected oocytes displayed two pronuclei 16 h post-insemination, while the proportion of embryos at 44-46 h post-insemination was 90.2%. At this time point, the majority (68.3%) of embryos were at the two-cell stage, showing in most cases (78.7%) minimal or moderate fragmentation. Eighteen clinical pregnancies, three of which were twin, were observed, giving rise to rates of 12.3 and 9.7%, calculated per patient and per embryo transfer, respectively.The implantation rate was 5.2%. To date, four children have been born and three pregnancies resulted in spontaneous abortions, while the remaining pregnancies are ongoing. CONCLUSIONS: Our data indicate that although the combination of slow cooling and high sucrose concentration ensures high rates of oocyte survival, it is not sufficient to guarantee a high standard of clinical efficiency.

Keywords: cryopreservation/fertility preservation/oocytes/pregnancy rate/slow cooling.
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