The use of Percoll gradients for the preparation of subpopulations of human spermatozoa

Abstract

Centrifugation of human spermatozoa on Biggers-Whitten-Whittingham or HEPES-Krebs-Ringer-buffered media containing Percoll as a method to prepare pure gamete populations or subpopulations was studied in detail. Application of this technique allowed uncontaminated, motile spermatozoa to be obtained and, after a subsequent Percoll gradient step, resulted in the enrichment of motile cells able to readily penetrate in vitro into AB-serum filled capillaries. The gradient sedimentation patterns of 25 different sperm samples with normal seminal parameters were strongly influenced by the characteristics of the sample itself and also by the buffer used for filtration. Ultrastructural changes and the release of intraspermatozoal enzymes were induced by the procedure. Although of interest clinically, the Percoll filtration technique in its present form is probably of limited value for cell biological and/or ultrastructural studies.