Sahabeh Etebary - Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Nahid Yari - Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Mohammad ali Khalili - Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Seyed Mehdi Kalantar - Reproductive and Genetic Unit, Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Bouali Ave, Safaeyeh, Yazd, Iran

Background: There are difficulties associated with testicular sperm freezing. Different methods of sperm cryopreservation were developed. Not enough detailed studies about the real efficacy of these techniques exist. For sperm morphologic assessment, MSOME is able to identify not only conventional morphological sperm alterations, but also sperm head vacuoles.Methods: After preparation, testicular sperm extraction samples of 15 azoospermic men, aged 20-40 years old were divided into three groups. Group 1 was assessed freshly. Group 2 was cryopreserved with vitrification method and Group 3 with cooling in liquid nitrogen vapor using droplet. Pre and post warming assessment in terms of spermatozoa head vacuoles by MSOME, DNA fragmentation and mitochondrial membrane potential was performed.Result: The number of spermatozoa with no vacuoles significantly decreased after two cryopreservation techniques (P < 0.001). There were no significant differences between groups regarding small and large vacuoles (p > 0.05). DNA fragmentation and mitochondrial membrane potential increased after cryopreservation (P < 0.001). There was a significant positive correlation between spermatozoa with large vacuoles in group 2 and DNA fragmentation and a significant positive correlation between spermatozoa with no vacuole and mitochondrial membrane potential in groups 2 and 3.Conclusion: Cryopreservation affects spermatozoa vacuolization, DNA structure and mitochondrial membrane potential. Using MSOME in selection of post thaw morphologically normal testicular spermatozoa for ICSI procedure will be of particular value.

Azoospermia, Cryopreservation, DNA Fragmentation, MSOME, Vacuoles, Spermatozoa