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Human Reproduction, Vol. 5, No. 1, pp. 109-115, 1990
© 1990 European Society of Human Reproduction and Embryology

research-article

Observations on the morphology of pronuclei and nucleoli in human zygotes and implications for cryopreservation

Graham Wright, Sharon Wiker, Carlene Elsner, Hilton Kort, Joe Massey, Dorothy Mitchell1, Andy Toledo1 and Jacques Cohen1,2

Reproductive Biology Associates Suite 330-D, 993 Johnson Ferry Road, Atlanta, GA 30342 1Gamete and Embryo Research Laboratory, Department of Gynecology and Obstetrics, Emory University Medical School Atlanta, GA, USA

Correspondence: 2To whom correspondence should be addressed at: Center for Reproductive Medicine and Infertility, Cornell University Medical College, Helmsley Medical Tower, Box l-HT-306, 505 East 70 Street, New York, NY 10021, USA

The effects of cryopreservation on human zygotes at various stages between the appearance of pronuclei and their close association were investigated. Pronuclear zygotes (n = 233) from 101 patients were frozen using propanediol 21–35 h following egg collection. The incidence of implantation of thawed pronuclear zygotes frozen 29–35 h following oocyte collection was significantly higher than that of younger pronuclear zygotes (28 versus 10%, respectively). Zygote age did not affect cell survival following cryostorage. The diameter and association of pronuclei and the number and distribution of nucleoli were determined from video tape recordings of 140 fresh zygotes. Pronuclear migration continued after pronuclear enlargement. The number of nucieoli remained constant during pronuclear migration, but their random distibution within the pronucleus diminished. Strongly adhered pronuclei had significantly more aligned nucleoli on adjacent sides than pronuclei which were still visually separated by ooplasm. This equatorial distribution of nucleoli was noted in the majority of zygotes older than 26 h. The findings suggest that zygote cryopreservation should be initiated when pronuclear migration is completed. This moment can be determined accurately by studying pronuclear association and nucleolar alignment.

Key words: pronucleus/nucleolus


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