P-150: Effect of Vitrification on High Magnification Morphology and Fertility Potential of Human Spermatozoa Cell

Background: Many sperm characteristics including, motility and morphology are important for normal spermatozoa- oocyte intraction. These features may be altered during vitrification. Motile sperm organelle morphology examination (MSOME) is an unstained real time high magnification technique that analysis viable sperm morphology instantly. This study aimed to investigate the influence of vitrification on human sperm vacuolization by MSOME technique and fertility potential by zona binding assay (ZBA). Materials and Methods: 30 normozoospermic ejaculates were prepared by direct swim up technique. After swim up, the supernatants were divided into two parts: fresh and vitrified groups. For vitrification, sperm suspension was mixed with equal volume (1:1) of Hams' F10+HSA+0.5 M sucrose, and 30 μl sperm suspension was dropped directly to LN. Warming was performed by quick submerging spheres one by one into pre-warmed 5ml Hams' F10 with 5% HSA at 37°C accompanied by gentel vortexing for 5-10s. Sperm motility and MSOME were evaluated for each sample.for MSOME, we loaded 2μl sperm suspension into 4 μl polyvinylpyrrolidone(PVP) in different culture dishes (GWST 5040; Will Co, Wells BV) and covered with oil.2×106 spermatozoa in each 25μl droplet containing four oocytes were performed for ZBA. Results: Our formula is (Normal head score=2)+ (Lack of vacuole score=3)+ (Normal base score=1)= (Total score=6) for a morphologic ‘‘normal top’’ spermatozoon.For MSOME analysis, 3 grades were considered: normal spermatozoa with no vacuoles (grade1), ≤2 small vacuoles (grade2), and at least one large vacuole or >2 small vacuoles and morphometrically abnormal spermatozoa (grade3). There was no significant difference between the rates of MSOME in class1 (1 ± 2.7), class2 (-0.5 ± 3.1), class3 (-1.11 ± 3.6) and fertility potential before and after vitrification Conclusion: Vitrification has adverse effect on sperm parameters. This technique, however, neither causes vacuolization of sperm head nor alters fertility potential.