Received July 19, 2022
Accepted December 02, 2022

Objective: Annexin V columns has been proposed as a tool to reduce levels of sperm apoptosis. The aim of this study was to determine the presence of nuclear vacuoles and apoptotic markers after using Annexin V columns and evaluate the size and number of nuclear vacuoles with sperm apoptosis. Methods: Double-blind prospective cohort study Ten semen samples, with a previously TUNEL assay (>20%), were analyzed. After two layers gradient, the pellet was eluted through Annexin V column. Three sperm populations were studied: after gradient, eluted fraction and the fraction retained in the column; evaluating the number and size of vacuoles, phosphatidylserine externalization, cleaved caspase-3 and TUNEL. Results: A significant decrease of apoptotic sperm markers levels was observed after using Annexin V columns (eluted fraction) (p<0.05). However, the numberand size of nuclear vacuoles do not showed a significant difference between fractions. The fraction with high levels of sperm apoptosis (fraction retained in the column) do not showed a prevalence of vacuolated spermatozoa and larger size of vacuoles. Conclusions: The sperm apoptosis levels significantly decreased after using Annexin V Columns, but not in the same way in relation to the number of vacuolated spermatozoa. There is not a correlation between a major number and size of vacuoles with high levels of sperm apoptosis.