Aryl Phosphate Derivatives of Bromo-Methoxy-Azidothymidine: MATERIALS AND METHODS(12)
Heterologous Sperm-Egg Binding and Penetration Assay
The fertilizing capacity of human sperm exposed to WHI-05 and WHI-07 was evaluated using the zona-free hamster egg penetration assay, with modifications. Motile fractions of sperm (n = 4) obtained by 90-45% Percoll gradient centrifugation and wash were resuspended in BWW-3.5% BSA medium (pH 7.4) and divided into aliquots of 20 X 106 sperm/ml and allowed to capacitate for 24 h. Subsequently, sperm were treated at 37°C for 3 h with 25 ^M or 100 ^M concentrations of WHI-05 and WHI-07 (in 0.5% DMSO) under capacitating conditions. After exposure to drugs, the control and WHI-treated sperm suspensions were washed in BWW-3.5% BSA medium. Sperm motility following capacitation and washing in BWW-3.5% BSA medium was assessed by CASA. buy ortho tri-cyclen
Frozen hamster eggs (Charles River Laboratories, Wilmington, MA) were thawed and transferred to 35-mm glass-bottom microwells (Mat Tek Corporation, Ashland, MA) in BWW medium. To remove zona, the washed eggs were treated with 0.05% trypsin (Irvine Scientific) in BWW medium for 20 min. Then 15 to 45 zona-free eggs per test were transferred into 100 ^l of BWW-3.5% BSA medium under mineral oil.