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Human Reproduction, Vol. 14, No. 2, 470-475, February 1999
© 1999 European Society of Human Reproduction and Embryology

Rapid visualization of metaphase chromosomes in single human blastomeres after fusion with in-vitro matured bovine eggs

Steen Willadsen1, Jacob Levron2, Santiago Munné1, Tim Schimmel1, Carmen Márquez1, Richard Scott1 and Jacques Cohen1,3

1 The Institute for Reproductive Medicine and Science of Saint Barnabas, Livingston, New Jersey, USA and 2 Department of Obstetrics and Gynecology, Tel Hashomer, Tel-Aviv, Israel

The present study was aimed to facilitate karyotyping of human blastomeres using the metaphase-inducing factors present in unfertilized eggs. A rapid technique for karyotyping would have wide application in the field of preimplantation genetic diagnosis. When cryopreserved in-vitro matured bovine oocytes were fused with human blastomeres, the transferred human nuclei were forced into metaphase within a few hours. Eighty-seven human blastomeres from abnormal or arrested embryos were fused with bovine oocytes in a preclinical study. Fusion efficiency was 100%. In 21 of the hybrid cells, no trace of human chromatin was found. Of the remaining 66, 64 (97%) yielded chromosomes suitable for analysis. The method was used to karyotype embryos from two patients with maternal translocations. One embryo which was judged to be karyotypically normal was replaced in the first patient, resulting in one pregnancy with a normal fetus. None of the second patient's embryos was diagnosed as normal, and hence none was transferred. The results of the present study demonstrated that the ooplasmic factors which induce and maintain metaphase in bovine oocytes can force transferred human blastomere nuclei into premature metaphase, providing the basis for a rapid method of karyotyping blastomeres from preimplantation embryos and, by implication, cells from other sources.

Key words: electrofusion/FISH/nuclear transplantation/spectral karyotyping/translocation

3 To whom correspondence should be addressed at: Institute for Reproductive Medicine and Science of Saint Barnabas, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052, USA


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