Human Reproduction, Vol. 3, No. 8, pp. 1010-1019, 1988
© 1988 European Society of Human Reproduction and Embryology
research-article |
Sexing whole human pre-embryos by in-situ hybridization with a Y-chromosome specific DNA probe
Department of Obstetrics and Gynaecology, University of Edinburgh, Centre for Reproductive Biology 37 Chalmers Street, Edinburgh EH3 9EW 1Medical Research Council, Clinical and Population Cytogenetics Unit, Western General Hospital Crewe Road, Edinburgh EH4 2XU, UK
Correspondence: 3To whom correspondence should be addressed
We have used DNA-DNA In-situ hybridization with a DNA probe for the human Y-chromosome to distinguish between male and female human pre-embryos. Both biotinylated and tritlated Y-probes worked well on control cell cultures where 100 interphase nuclei were scored. Morphologically normal pre-embryos could be sexed with confidence with the tritiated Y-probe but the biotin results were less reliable (although only a few pre-embryos were analysed in this way). Early cleavage stage pre-embryos had large nudel with relatively diffuse V-bodies and were more difficult to score with the biotinylated Y-probe. Morphologically abnormal pre-embryos often had large nuclei with multiple Y-hodies (presumably polyploid nuclei) or small nuclei with no Y-bodies (possibly fragmenting nudei). In all, 38 clenying and two non-cleaving pre-embryos were analysed. The incidence of false positive and false negative cells seen after hybridization of tritiated Y-probes to control lymphocyte cultures suggests that it should normally be possible to distinguish morphologically normal male and female pre-embryos with samples of three to six interphase nuclei.
Key words: Y-chromosome/DNA-DNA hybridization/sexing/embryos/human
2Present address: Department of Obstetrics and Gynaecology, Yale University School of Medicine, 340 Farnum Memorial Building, PO Box 3333, New Haven, CT 06510, USA