Effect of multiple centrifugations on the evaluation of the acrosoome reaction in human spermatozoa
This study was undertaken to assess the effect of multiple centrifugations on the human acrosome reaction and in vitro fertilizing ability. Semen samples were obtained from sperm donors and from patients participating in our in vitro fertilization programme. Measured sperm samples were centrifuged twice (400xg for 5 min) before or after the induction of the acrosome reaction with calcium ionophore A23187 and before the insemination of oocytes. The samples obtained from the sperm donors were used to measure the acrosomal loss by means of indirect immunofluorescence (T6 monoclonal antibody) and the patient samples were used to assess fertilizing ability and cleavage. Samples centrifuged twice before the chemical induction of the acrosome reaction exhibited significantly elevated levels of acrosomal loss (mean = 46%) as compared to the controls (23%). However, the fertilization (P = 0.688) and cleavage (P = 0.187) rates were not significantly different between the controls and the centrifuged samples. The present study has shown that the centrifugation of motility enriched (swim-up) samples may also modify the acrosomal membrane complex, without compromising fertilizing potential.