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Human Reproduction, Vol. 14, No. 2, 400-408, February 1999
© 1999 European Society of Human Reproduction and Embryology

Vitrification of mouse germinal vesicle oocytes: effect of treatment temperature and egg yolk on chromatin and spindle normality and cumulus integrity

E.F. Isachenko1 and P.L. Nayudu

Department of Reproductive Biology, German Primate Center, Kellnerweg 4, Göttingen, 37077, Germany

The success rates for cryopreservation of immature oocytes from several species including human remain low, in contrast to major improvements with mature oocytes. In this study, a new approach has been developed using a short exposure ultra-rapid freezing protocol, examining the effect of temperature and egg yolk (two factors which may be expected to influence membrane flexibility) on the cryostability of immature mouse oocytes and cumulus complexes. These two factors were tested in various patterns for their cryoprotective effect using ethylene glycol as the principal cryoprotectant. The results showed that 37°C pre- and post-freeze exposure significantly improved both survival and normal spindle configuration after in-vitro maturation. Egg yolk was found to produce further beneficial effects on both the oocyte and cumulus cell integrity, with the best effects being obtained at 37°C with inclusion of egg yolk both before and after the freezing. This protocol produced >80% normal survival post-thaw with intact and attached cumulus complex, 84% maturation rate and 45% normal metaphase configuration. In summary, a unique combination of high survival and meiotic normality together with good preservation of the attached cumulus cell mass has been achieved using a simple new vitrification procedure.

Key words: chromatin/germinal vesicle oocytes/mouse/rapid freezing/spindle

1 To whom correspondence should be addressed


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