Abstract
The aim of this study was to investigate the differences in chromatin stability of native and frozen–thawed spermatozoa of fertile and subfertile men. Forty fertile and 28 subfertile men were included in this study. A 1 ml of each native semen sample was mixed 1:1 with lithium diiodosalycylate (LIS)–dithiothreitol (DTT) for 60 min and 1 ml semen was mixed 1:1 with Human Sperm Protection Medium (HSPM) and frozen.After thawing, the cryoprotective was washed away and each sample was also mixed 1:1 with LIS–DTT for 60 min. Many smears were made directly after liquefaction as well as 30 and 60 min post-incubation from fresh and frozen–thawed spermatozoa. Human nuclear chromatin decondensation (NCD) was evaluated at different time intervals using IBAS. Incubation of spermatozoa with LIS+DTT led to sperm area enlargement (swelling) in both groups. However, by incubation of spermatozoa after freeze-thawing, the area enlargement increased significantly ( p=0.001), in both groups in comparison to native semen samples. Besides, the area enlargement of frozen–thawed spermatozoa differs significantly ( p=0.001) between the two groups after 60 min incubation. In conclusion: decondensation of spermatozoa led to a significant difference between the area before and after the freeze-thawing process. However, spermatozoa from fertile group are more condensed and stabile than those of subfertile spermatozoa.
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