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- Title
Effect of vitrification on high magnification morphology, chromatin condensation, and fertility potential of human spermatozoa.
- Authors
Taherzadeh S.; Khalili M. A.; Anvari M.; Ghazali Sh.; Agha-Rahimi A.; Anbari F.; Ghasemi-Esmailabad S.
- Abstract
Introduction: Sperm vitrification is a technique of iceand CPA- free cryopreservation by direct plunging of a sperm suspension into liquid nitrogen (LN). Sperm characteristics of motility and morphology are important for normal spermatozoa-oocyte interaction. However, defects on chromatin condensation can cause vacuolization on sperm morphology. Motile sperm organelle morphology examination (MSOME) is an unstained real time high magnification that analysis viable sperm morphology. This study investigated the influence of vitrification on human sperm structure by MSOME technique, and fertility potential by zona binding assay (ZBA) and chromatin condensation by toluidine blue (TB) and aniline blue (AB) assessment. Materials and Methods: 30 normozoospermic ejaculates were prepared by swim up technique, and supernatants were divided into two parts: fresh and vitrified groups. For vitrification, sperm suspension was mixed with equal volume (1:1) of Hams F10 +5% HSA +0.5 M sucrose. Then, 30 pi sperm suspension was dropped into LN. Warming was performed by quick submerging spheres into pre-warmed 5ml Hams F10 with 5% HSA at 37°C. Sperm motility, stained morphology, MSOME and ZBA were evaluated for each sample. Three classes were considered for MSOME analysis: high quality sperm with a score of 4- 6 (Class 1); medium-quality sperm with a score of 1-3 (Class 2); low-quality sperm with a score of 0 (Class 3). 2×106 spermatozoa in each 25pi droplet containing 4 Oocytes were performed for ZBA. In addition, samples were fixed for TB and AB assessments. Results: Cryopreservation significantly reduced both progressive motility and normal morphology of spermatozoa. There was no significant differences between the rates of MSOME in class 1 (14.93±14.66 vs. 13.56±11.34), class 2 (53.53±13.99 vs. 55.63±12.16), class 3 (31±20.81 vs. 30.80±16.03) pre and post vitrification, respectively. However, vitrification reduced the fertility potential of spermatozoa from normozoospermic samples (13.40±22.73 vs. 9.00±13.87) and chromatin condensation (TB: 60.32±16.60 vs. 57.28±17.19) (AB: 38.32±8.14 vs. 34.35±6.87). Conclusion: Vitrification had adverse effects on sperm parameters of motility and morphology. However, this technique did not increase the rate of vacuolization of sperm head or severe alteration in fertility potential. Since, the majority of human spermatozoa contained vacuolization in head region, it is highly recommended to use MSOME technology for assessment of sperm fine morphology during clinical microinjection procedure.
- Subjects
SPERMATOZOA; CHROMATIN; CRYOPRESERVATION of organs, tissues, etc.
- Publication
Iranian Journal of Reproductive Medicine, 2015, p81
- ISSN
1680-6433
- Publication type
Abstract