Karyotyping of human oocytes by cenM-FISH, a new 24-colour centromere-specific technique.
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.
Gutiérrez-Mateo
C
C
Departament de Biologia Cellular, Fisiologia i Immunologia, Unitat de Biologia Cellular i Genètica Mèdica, Facultat de Medicina, Universitat Autònoma de Barcelona, E-08193 Bellaterra, Spain. cristina.gutierrez@uab.es
Benet
J
J
Starke
H
H
Oliver-Bonet
M
M
Munné
S
S
Liehr
T
T
Navarro
J
J
D000328:Adult; D000373:Agglutinins / Q000378:metabolism; D000782:Aneuploidy; D002503:Centromere / Q000648:ultrastructure*; D002869:Chromosome Aberrations; D002877:Chromosomes, Human / Q000648:ultrastructure*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007247:Infertility, Female; D007621:Karyotyping / Q000379:methods*; D027806:Lens Plant; D008677:Metaphase; D009693:Nucleic Acid Hybridization; D009865:Oocytes / Q000166:cytology / Q000378:metabolism
D000373:Agglutinins; D005456:Fluorescent Dyes