Human spermatozoa selection in improved discontinuous Percoll gradients.
OBJECTIVE
Improve the technique of human spermatozoa separation using Percoll density gradients.


METHODS
We compared the spermatozoa separation obtained after density gradients from a physiological stock Percoll solution and a hyperosmotic stock Percoll solution.


METHODS
Sperm samples were obtained from the Laboratory of the Biology of Reproduction at Brest, France, a public hospital.


METHODS
One hundred thirty-eight healthy donors.


METHODS
None.


METHODS
The resolution of the separation was assessed by the number of motile spermatozoa recovered after separation using density gradients.


RESULTS
The improvement of the resolution of separation is at least twofold greater with the use of hyperosmotic gradients than with that obtained by physiological gradients. Moreover, the best resolutions of separation were obtained from pathological sperm. Finally, the influence of buoyant density spermatozoa on the resolution of separation by this technique is clearly demonstrated.


CONCLUSIONS
With this modification, the technique of Percoll density gradients, used for the separation of human spermatozoa, is strongly advised especially in pathological cases.
Laboratoire de Cytogénétique, Cytologie et Biologie de la Reproduction, CHU de Brest, France.
J F
JF
Velez de la Calle
D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D006801:Humans; D008297:Male; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology