title: Comparison of two techniques to evaluate the acrosomal status of zona pellucida bound sperm in humans.
abstract: In this work, we have compared two procedures that evaluate the acrosomal status of human sperm bound to the human zona pellucida. Motile sperm, selected by a Percoll gradient, were capacitated by incubation at 37 degrees C, 5% CO2, for 4.5 h, at 20 x 10(6) cells ml(-1). Then, the sperm were incubated with nonviable human oocytes for 10 min at 37 degrees C, 5% CO2. The oocytes with bound sperm were transferred to 500 microl phosphate-buffered saline (PBS) and washed to remove loosely bound sperm. The oocytes were then processed according to the procedures of Cross et al. (1986) or Liu & Baker (1996). In the Cross's procedure, the sperm were labelled while they were bound to the zona. In the Liu's procedure, the sperm were first dislodged from the zona into a droplet of PBS and labelled in there. Both procedures gave equivalent percentages of acrosome-reacted sperm. However, the total number of zona-bound sperm available for assessment with the procedure of Liu & Baker was greater than that of Cross et al. We suggest to use the former procedure to evaluate the acrosomal status of zona-bound sperm in humans. Moreover, this procedure also provided information about sperm ability to bind to the zona pellucida.
authors.affiliation: Department of Biomedicine, Faculty of Health Sciences, University of Antofagasta, Antofagasta, Chile. pmorales@uantof.cl
authors.forename: P
authors.initials: P
authors.lastname: Morales
authors.forename: C
authors.initials: C
authors.lastname: Pasten
mesh_terms: D020101:Acrosome Reaction; D033441:Andrology / Q000379:methods*; D006690:Bisbenzimidazole; D002102:Cadaver; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D013094:Spermatozoa / Q000502:physiology*; D015044:Zona Pellucida / Q000502:physiology*
chemical_list: D005456:Fluorescent Dyes; D006690:Bisbenzimidazole