High quality human sperm selection for IVF: A study on sperm chromatin condensation

The study consisted of semen samples of 20 male individuals who applied to Abant Izzet Baysal University Faculty of Medicine and participated in a spermiogram. The aim of this study was to determine how to obtain the healthiest spermatozoa by employing a variety of swim-up methods over differing time periods and without the use of centrifuge. Ejaculate samples were taken from the 20 patients and each patient's homogenized semen sample was divided into 4 groups without centrifugation. Group 1 was taken as the sample of untreated semen. For the other 3 groups, 250 mu l of medium was added in the semen samples. Afterwards, the samples were kept at 37 degrees C for different time periods, 30 min for Group 2, 60 min for Group 3 and 90 min for Group 4 in order for the spermatozoa to swim to the media in the upper layer. At the end of the periods, 10 mu l of propagation preparations were prepared from the swim-up fluid. Using Aniline Blue for chromatin condensation analysis, two hundred cells were immunostained by Caspase 3 for apoptotic analysis. Subsequently, the result of the four groups were compared for each test. The spermatozoa obtained at the end of the 30 min. of swim-up was compared to the spermatozoa obtained from the swim-up of 60 min., the swim-up of 90 min. It was found that the control group had statistically significant lower rates of apoptosis and was healthier in terms of chromatin integrity. The swim-up method without centrifugation is the best suited sperm preparation, based on sperm DNA integrity and sperm chromatin condensation.