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Hum. Reprod. Advance Access published online on August 26, 2005
Human Reproduction, doi:10.1093/humrep/dei252
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
Received April 15, 2005
Revised July 12, 2005
Accepted July 14, 2005

Article

Karyotyping of human oocytes by cenM-FISH, a new 24-colour centromere-specific technique

C. Gutiérrez-Mateo 1*, J. Benet 1, H. Starke 2, M. Oliver-Bonet 1, S. Munné 3, T. Liehr 2, and J. Navarro 1*

1 Departament de Biologia Cel·lular, Fisiologia i Immunologia, Unitat de Biologia Cel·lular i Genètica Mèdica, Facultat de Medicina, Universitat Autònoma de Barcelona, E-08193 Bellaterra, Spain
2 Institute of Human Genetics and Anthropology, 07740 Jena, Germany
3 The Institute for Reproductive Medicine and Science, St Barnabas Medical Center, 94 Old Short Hills Road, Livingston, NJ 07039, USA; Reprogenetics, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052, USA

* To whom correspondence should be addressed.
C. Gutiérrez-Mateo, E-mail: cristina.gutierrez{at}uab.es
J. Navarro, E-mail: joaquima.navarro{at}uab.es


  Abstract

BACKGROUND: Metaphase II (MII) chromosome complements are difficult to karyotype. The objective of this study was to investigate the efficiency and limitations of centromere-specific multiplex fluorescence in situ hybridization (cenM-FISH), a new 24 colour FISH technique using centromere-specific probes, to analyse the whole chromosome complement within human oocytes. METHODS: Oocytes were donated by 34 patients undergoing ovarian stimulation and IVF. The MII oocytes were analysed by means of cenM-FISH, while the confirmation of results was performed by FISH and/or by analysing the corresponding first polar bodies using comparative genomic hybridization (CGH). RESULTS: A total of 30 cells, corresponding to 16 oocytes and 14 first polar bodies, were successfully karyotyped by either cenM-FISH or CGH. The incidence of aneuploidy was 25%, and eight out of nine aneuploidy events were confirmed by CGH and FISH. CONCLUSIONS: We demonstrate here for the first time that the identification of any numerical abnormality in oocytes is feasible using cenM-FISH. Despite the fact that the fixation efficiency remains low, the present results confirm the advantage of analysing the whole set of chromosomes to make an accurate estimation of the aneuploidy rate in human oocytes.

Keywords: aneuploidy/cenM-FISH/CGH/first polar body/oocyte.
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