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Hum. Reprod. Advance Access published online on May 16, 2006
Human Reproduction, doi:10.1093/humrep/del157
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received February 14, 2006
Revised March 25, 2006
Accepted April 11, 2006

Article

Comparative genomic hybridization analysis of human oocytes and polar bodies

E. Fragouli 1 *, D. Wells 2, A. Thornhill 3, P. Serhal 4, M.J.W. Faed 5, J.C. Harper 1, and J.D.A. Delhanty 1

1 Department of Obstetrics and Gynaecology, UCL Centre for Preimplantation Genetic Diagnosis, University College London, London, UK
2 Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, CT, USA
3 The London Fertility Centre, University College London Hospitals, Eastman Dental Hospital, London
4 The Assisted Conception Unit, University College London Hospitals, Eastman Dental Hospital, London
5 Department of Molecular and Cellular Pathology, Ninewells Hospital and Medical School, University of Dundee, Dundee, Scotland, UK

* To whom correspondence should be addressed.
E. Fragouli, E-mail: efragouli{at}hotmail.com


  Abstract

BACKGROUND: Classical cytogenetic methods and fluorescent in situ hybridization (FISH) have been employed for the analysis of chromosomal abnormalities in human oocytes. However, these methods are limited by the need to spread the sample on a microscope slide, a process that risks artefactual chromosome loss. Comparative genomic hybridization (CGH) is a DNA-based method that enables the investigation of the entire chromosome complement. We optimized and evaluated a CGH protocol for the chromosomal analysis of first polar bodies (PBs) and oocytes. The protocol was then employed to obtain a detailed picture of meiosis I errors in human oogenesis. METHODS: 107 MII oocyte-PB complexes were examined using whole genome amplification (WGA) and CGH. RESULTS: Data was obtained for 100 complexes, donated from 46 patients of average age 32.5 (range 18-42). 22 complexes from 15 patients were abnormal, giving an aneuploidy rate of 22%. CONCLUSIONS: The results presented in this study more than double the quantity of CGH data from female gametes currently available. Abnormalities caused by whole chromosome non-disjunction, unbalanced chromatid predivision and chromosome breakage were reliably identified using the CGH protocol. Analysis of the data revealed a preferential participation of chromosome X and the smaller auto-somes in aneuploidy and provided further evidence for the existence of age-independent factors in female aneuploidy.

Keywords: aneuploidy/chromosome abnormalities/comparative genomic hybridization/oocyte/polar body.
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