A new method of the electrolyte-free long-term preservation of human sperm at 4 degrees C
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K Saito1, Y Kinoshita, H Kanno
1Department of Urology, Yokohama City University School of Medicine, Japan.
Objectives:
To develop a new method for the long-term preservation of human sperm.
Setting:
Andrology laboratory of our hospital.
Patients:
Thirty-one normal and 19 asthenozoospermic semen samples obtained from patients attending our infertility clinic. The average sperm motility was 70.2% and 36.0% in the normal and asthenozoospermic groups, respectively.
Interventions:
Ejaculated sperm were centrifuged and washed using the electrolyte-free Percoll gradient and then were preserved at 4 degrees C.
Main Outcome Measures:
The motility of the preserved sperm was analyzed using computer-assisted semen analyzer after the addition of Ham's modified F-10 (GIBCO, Grand Island, NY).
Results:
In the normal group, motility rate after the addition of Ham's F-10 for 1, 2, and 4 weeks of preservation was 65.4%, 40.4%, and 5.5%, respectively. In the asthenozoospermic group, motility rate after 1 and 2 weeks of preservation was 31.3% and 18.1%, respectively. Preservation solutions containing sodium or potassium decreased motility after preservation. Restoration of preserved sperm was not achieved by incubation alone; however, reinitiation was induced by incubation together with Ham's F-10.
Conclusions:
Human sperm in the electrolyte-free solution survived for a long period of time at 4 degrees C and reinitiation of sperm motility after preservation required the addition of Ham's F-10.