Nuclear DNA Normality of Spermatozoa Selected by Swim-up and Two-layer Percoll Gradient Techniques

Abstract

Objective To use acridine orange (AO) test to compare sperm DNA normality between washing and swim-up, and two-layer Percoll gradient techniques for both normal and abnormal semen samples.

Setting Division of Infertility, Department of Obstetrics and Gynaecology, Faculty of Medicine, Siriraj Hospital, Mahidol University.

Subjects and methods Semen samples from 112 male partners of infertile couples were divided into two aliquots, which were separated by washing and swim-up, and two-layer Percoll gradient techniques. AO test was performed on the sperm before and after sperm separation.

Results The percentage of green-fluorescing sperm is improved after the two methods of sperm preparation but is more improved after two-layer Percoll gradient than washing and swim-up , both in normal and abnormal semen samples (89.0 + 7.9 versus 81.9 + 10.2, and 85.2 + 10.2 versus 74.6 + 14.8; p < 0.001 respectively).

Conclusion Two-layer Percoll gradient selected more sperm with nuclear DNA normality than washing and swim-up separation that may influence the outcome of assisted reproductive techniques.