Assessment of the separation of X- and Y-bearing sperm on albumin gradients using double-label fluorescence in situ hybridization.
OBJECTIVE
To determine the ratio of X- to Y-bearing human spermatozoa in fractions isolated from discontinuous albumin gradients.


METHODS
The proportions of X- and Y-bearing sperm were determined in neat semen samples (control) and in albumin-separated fractions from the same samples. Two albumin methods were used: a two-layer method (experiment 1) and a three-layer method (experiment 2). X- and Y-bearing sperm were identified simultaneously using chromosome-specific DNA probes and fluorescence in situ hybridization.


METHODS
Hospital-based university department.


METHODS
Healthy donors with normal semen characteristics.


METHODS
The proportions of haploid cells (X or Y) and cells with two sex chromosomes (XX, YY, or XY) were determined.


RESULTS
Labeling efficiencies were > 96% in all samples. Control samples showed a 1:1 ratio of X- to Y-bearing sperm. Fractions isolated on albumin gradients showed a slight, but statistically significant enrichment of X-bearing sperm. This was evident with both albumin methods.


CONCLUSIONS
Discontinuous albumin gradients do not enrich Y-bearing sperm as previously reported.
Wang
H X
HX
Department of Obstetrics and Gynaecology, University of Adelaide, Queen Elizabeth Hospital, Woodville, South Australia.
Flaherty
S P
SP
Swann
N J
NJ
Matthews
C D
CD
D000782:Aneuploidy; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D015342:DNA Probes; D006238:Haploidy; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D012709:Serum Albumin; D013094:Spermatozoa / Q000648:ultrastructure*; D014960:X Chromosome; D014998:Y Chromosome
D015342:DNA Probes; D012709:Serum Albumin