Effect of vitrification on high magnification morphology of human spermatozoa cells

Taherzadeh, S.; Khalili, M. A.; Anvari, M.; Agha-Rahimi, A.; Ghazali, S.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p25
Academic Journal
Introduction: Human sperm vitrification is a new technique of cryoprotectant-free cryopreservation that has been developed recently. Motile sperm organelle morphology examination (MSOME) is a technique that analysis viable sperm morphology exactly. This study aimed to investigate the influence of vitrification on sperm vacuolization by MSOME technique. Materials and Methods: 30 normozoospermic ejaculated samples were obtained after at least 48h of sexual abstinence. Semen was prepared by direct swim-up technique. The prepared spermatozoa 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 ul sperm suspension was dropped directly to liquid nitrogen. Warming was performed by quickly submerging spheres one by one (not more than ten spheres) into 5ml Hams f10 with 5%HSA pre-warmed at 37°C accompanied by gentel vortexing for 5-10s. Sperm motility, viability & MSOME were evaluated for each sample. For MSOME, we loaded 2ul sperm suspension into 4 ul polyvinylpyrrolidone (PVP) in different culture dishes (GWST 5040; Will Co, Wells BV) and covered with oil. 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 (gradel), <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 classl (1±2.7), cla ss2 (-0.5±3.1), class3 (-1.11±3.6) before and after vitrification. Conclusion: Vitrification has adverse effect on sperm parameters, but this technique does not cause vacoulisation of head and does not increase the abnormal morphology of head.


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