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Human Reproduction, Vol. 17, No. 1, 178-183, January 2002
© 2002 European Society of Human Reproduction and Embryology

Germinal vesicle transfer between fresh and cryopreserved immature mouse oocytes

Federica Moffa1,2,*, Francesca Comoglio1,2,*, Lewis C. Krey1, James A. Grifo1, Alberto Revelli2,3, Marco Massobrio2 and John Zhang1

1 Program for In vitro Fertilization, Reproductive Surgery and Infertility, NYU School of Medicine, New York, USA and 2 Department of Obstetrical and Gynaecological Sciences, University of Torino, S. Anna Hospital, Torino, Italy

BACKGROUND: We assessed the maturational competence and the chromosomal pattern of mouse oocytes reconstructed by germinal vesicle (GV) transfer technique using nuclear and/or cytoplasmic components from cryopreserved GV stage oocytes. METHODS: From 657 GV oocytes (326 fresh and 331 frozen/thawed), four groups of reconstructed oocytes were obtained by micromanipulation and electrofusion: fresh GV–fresh cytoplast (FF), thawed GV–thawed cytoplast (TT), fresh GV–thawed cytoplast (FT), thawed GV–fresh cytoplast (TF). All reconstructed oocytes were cultured in vitro to metaphase II. RESULTS: Survival rate after manipulation and electrofusion, as well as progression to metaphase II, did not differ significantly among the four groups. Comparing reconstructed oocytes with fresh and thawed control pools, the only difference was a slightly but significantly higher maturation rate in the TT pool versus matched controls (P < 0.01). Cytogenetic analysis of 25 reconstructed oocytes showed the expected number of 20 chromosomes in 88% of them. CONCLUSIONS: We conclude that both nuclear and cytoplasmic components derived from cryopreserved immature oocytes are suitable for GV transfer procedure, and generate chromosomally normal oocytes able to progress to metaphase II in vitro. The possibility of using cryostored immature oocytes as a source of nuclei and cytoplasm could help in applying GV transfer procedure, both in research and clinical settings.

Key words: cryopreservation/germinal vesicle transfer/in-vitro maturation/nuclear transfer/oocyte

3 To whom correspondence should be addressed at: Department of Obstetrical and Gynaecological Sciences, University of Torino, S. Anna Hospital, Via Ventimiglia 3, 10126 Torino, Italy. E-mail: fertisave{at}yahoo.com

* F.Moffa (e-mail: femoffa{at}hotmail.com) and F.Comoglio (e-mail: fcomoglio{at}yahoo.com) contributed equally to this work.


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L.-B. Cui, X.-Y. Huang, and F.-Z. Sun
Transfer of germinal vesicle to ooplasm of young mice could not rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice
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