Genetics: Discontinuous Percoll gradients enrich X-bearing human spermatozoa: a study using double-label fluorescence int-situ hybridization |
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Authors: | Wang, Huai-Xiu Flaherty, Sean P. Swann, Nicholas J. Matthews, Colin D. |
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Affiliation: | Department of Obstetrics and Gynaecology, The University of Adelaide, The Queen Elizabeth Hospital Woodville South Australia 5011, Australia |
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Abstract: | The aim of this study was to evaluate the efficacy of enrichingX-bearing human spermatozoa using 12-step (2580%) discontinuousPercoll gradients. X- and Y-bearing spermatozoa were simultaneouslyidentified in neat semen (controls) and in 80% Percoll fractionsfrom the same samples using double-label fluorescence in-situhybridization and chromosome-specific DNA probes. Hybridizationand labelling efficiencies of 9599% were obtained inall samples. The mean ratio of X- to Y-bearing spermatozoa inthe controls was 49.0: 48.2, whereas there was a significantenrichment (P < 0.0001) of X-bearing spermatozoa in the Percollfractions (mean X:Y ratio was 55.1:41.1 or 1.35). The ratiovaried from 1.11.5 in individual Percoll samples. Therewere no significant differences in the proportions of aneuploidspermatozoa (XX, YY, XY) between the control and Percoll fractions.We conclude that 12-step discontinuous Percoll gradients enrichX-bearing spermatozoa, but the degree of enrichment is insufficientfor use in preconceptional sex selection. |
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Keywords: | human spermatozoa/fluorescence in-situ hybridization/Percoll/sex selection |
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