Human Reproduction, Vol. 15, No. 8, 1787-1790,
August 2000
© 2000 European Society of Human Reproduction and Embryology
Ultra-rapid freezing of human multipronuclear zygotes using electron microscope grids
1 Maria Infertility Medical Institute, 2 Department of Animal Sciences, Kon-Kuk University and 3 Maria Infertility Clinic, Seoul, Korea
Developmental capacity of human multipronuclear (PN) zygotes cryopreserved using an ultra-rapid freezing method and electron microscope (EM) grids was studied. Multipronuclear zygotes obtained from a human IVF programme were used as an alternative to normal 2PN zygotes; they were divided into 3PN or 
4PN zygotes and their in-vitro development and cryo-injury were compared according to PN number. EFS30, which consisted of 30% ethylene glycol, 18% Ficoll, 0.5 mol/l sucrose and 10% fetal bovine serum with added modified Dulbecco's phosphate buffered saline was used as the freezing solution. After ultra-rapid freezing and thawing 85.5% of multipronuclear zygotes survived. A comparison of cleavage rates between the control and cryopreserved groups showed no significant difference (3PN; 81.3 and 85.4% and 4PN; 90.0 and 95.7% respectively). Comparing the in-vitro development after thawing up to blastocyst formation on day 5 after IVF, the outcome of the frozen 3PN group (22.0%) was not different from that of control 3PN group (38.5%), while the outcome of the frozen 4PN group (4.5%) was significantly lower than that of control 4PN group (44.4%) (P < 0.05).
Key words: developmental capacity/EM grid/human/multipronuclear zygotes/ultra-rapid freezing
4 To whom correspondence should be addressed at: Maria Infertility Medical Institute, 103–11 Sinseol-dong Dongdaemun-Gu, Seoul 130–110, Korea. E-mail: mimi60{at}unitel.co.kr
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