The heparin-glutathione test: an alternative to the hypo-osmotic swelling test to select viable sperm for intracytoplasmic sperm injection

Abstract

Objective: To evaluate the heparin-glutathione test (HEGLUT) for the selection of viable sperm for intracytoplasmic sperm injection (ICSI). Design: A prospective study. Setting: Department of Pediatrics, Obstetrics and Gynecology, University of Valencia and Instituto Valenciano de Infertilidad. Patient(s): Semen samples from healthy donors and patients with infertility. Intervention(s): Sperm samples were kept in culture for different periods in Ham’s F-10 medium supplemented or not supplemented with heparin, reduced glutathione (GSH), or a heparin-GSH mixture. Control and heparin-GSH–treated spermatozoa were injected into hamster oocytes. The HEGLUT and ICSI were performed. Main Outcome Measure(s): Sperm nuclear decondensation, progressive and nonprogressive motility, and male pronucleus formation. Result(s): The maximum proportion of sperm nuclear decondensation (28.7% ± 2.1% versus 2.6% ± 0.5% in the control group) was reached after 60 minutes of incubation in the presence of a heparin-GSH mixture. Differences in the percentages of progressive and nonprogressive motility among treatments and times of incubation, although statistically significant, were biologically negligible. No statistically significant differences were observed in the rate of sperm head decondensation (8.2% [4/49] versus 11.1% [6/54]) and male pronucleus formation (18.4% [9/49] versus 22.2% [12/54]) after the injection of control and treated spermatozoa into hamster oocytes. Conclusion(s): The HEGLUT may offer an alternative to the hypo-osmotic swelling test for the selection of viable sperm for ICSI.