Abstract
Purpose
To evaluate whether sperm preparation (swim-up technique) before freezing improves the percentages of sperm motility, sperm viability, and non-apoptotic spermatozoa after freezing-thawing process compared with preparation after cryopreservation.
Methods
Semen samples from 65 infertile males were equally divided into two aliquots one of which was processed for swim-up prior to cryopreservation and one of which was processed following cryopreservation. Sperm count, motility, and apoptosis index were measured in each group.
Result (s)
The total sperm count and the total motile sperm count decreased after thawing in both the pre-preparation and non-preparation groups compared with neat semen group (P < 0.001). Moreover, the percentage of apoptotic sperm in the pre-preparation group after cryopreservation was lower than that in the non-preparation group (P < 0.05), whereas the percentage of vital sperm with progressive motility was higher than that in the pre-preparation group (P < 0.001).
Conclusion (s)
Semen preparation by swim-up before freezing resulted in better sperm quality and fewer apoptotic sperm than sperm preparation after thawing. Therefore, sperm preparation before cryopreservation should be considered in routine sperm cryopreservation.
Similar content being viewed by others
References
Nangia AK, Krieg SA, Kim SS. Clinical guidelines for sperm cryopreservation in cancer patients. Fertil Steril. 2013;100:1203–9.
Petyim S, Choavaratana R. Cryodamage on sperm chromatin according to different freezing methods, assessed by AO test. J Med Assoc Thai. 2006;89:306–13.
Critser JK, Arneson BW, Aaker DV, Huse-Benda AR, Ball GD. Cryopreservation of human spermatozoa. II. Postthaw chronology of motility and of zona-free hamster ova penetration. Fertil Steril. 1987;47:980–4.
Hammadeh ME, Askari AS, Georg T, Rosenbaum P, Schmidt W. Effect of freeze-thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men. Int J Androl. 1999;22:155–62.
Royere D, Hamamah S, Nicolle JC, Lansac J. Chromatin alterations induced by freeze-thawing influence the fertilizing ability of human sperm. Int J Androl. 1991;14:328–32.
Thomson LK, Fleming SD, Aitken RJ, De Iuliis GN, Zieschang JA, Clark AM. Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis. Hum Reprod. 2009;24:2061–70.
Abush A, Hauser R, Paz G, Kleiman SE, Lehavi O, Yavetz H, et al. Thawed human sperm quality is influenced by the volume of the cryopreserved specimen. Fertil Steril. 2014;101:640–6.
Ribas-Maynou J, Fernandez-Encinas A, Garcia-Peiro A, Prada E, Abad C, Amengual MJ, et al. Human semen cryopreservation: a sperm DNA fragmentation study with alkaline and neutral Comet assay. Andrology. 2014;2:83–7.
Larson KL, DeJonge CJ, Barnes AM, Jost LK, Evenson DP. Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Hum Reprod. 2000;15:1717–22.
Hoshi K, Katayose H, Yanagida K, Kimura Y, Sato A. The relationship between acridine orange fluorescence of sperm nuclei and the fertilizing ability of human sperm. Fertil Steril. 1996;66:634–9.
Twigg J, Fulton N, Gomez E, Irvine DS, Aitken RJ. Analysis of the impact of intracellular reactive oxygen species generation on the structural and functional integrity of human spermatozoa: lipid peroxidation, DNA fragmentation and effectiveness of antioxidants. Hum Reprod. 1998;13:1429–36.
Ricci G, Perticarari S, Boscolo R, Simeone R, Martinelli M, Fischer-Tamaro L, et al. Leukocytospermia and sperm preparation–a flow cytometric study. Reprod Biol Endocrinol. 2009;7:128.
Demir B, Arikan II, Bozdag G, Esinler I, Karakoc Sokmensuer L, Gunalp S. Effect of sperm morphology on clinical outcome parameters in ICSI cycles. Clin Exp Obstet Gynecol. 2012;39:144–6.
Whittington K, Ford WC. Relative contribution of leukocytes and of spermatozoa to reactive oxygen species production in human sperm suspensions. Int J Androl. 1999;22:229–35.
Aitken RJ, Buckingham D, West K, Wu FC, Zikopoulos K, Richardson DW. Differential contribution of leucocytes and spermatozoa to the generation of reactive oxygen species in the ejaculates of oligozoospermic patients and fertile donors. J Reprod Fertil. 1992;94:451–62.
Henkel R, Kierspel E, Stalf T, Mehnert C, Menkveld R, Tinneberg HR, et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril. 2005;83:635–42.
Said TM, Agarwal A, Sharma RK, Mascha E, Sikka SC, Thomas Jr AJ. Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility. Fertil Steril. 2004;82:871–7.
Brugnon F, Ouchchane L, Pons-Rejraji H, Artonne C, Farigoule M, Janny L. Density gradient centrifugation prior to cryopreservation and hypotaurine supplementation improve post-thaw quality of sperm from infertile men with oligoasthenoteratozoospermia. Hum Reprod. 2013;28:2045–57.
Valcarce DG, Carton-Garcia F, Riesco MF, Herraez MP, Robles V. Analysis of DNA damage after human sperm cryopreservation in genes crucial for fertilization and early embryo development. Andrology. 2013;1:723–30.
Rougier N, Uriondo H, Papier S, Checa MA, Sueldo C, Alvarez SC. Changes in DNA fragmentation during sperm preparation for intracytoplasmic sperm injection over time. Fertil Steril. 2013;100:69–74.
Justice T, Christensen G. Sperm cryopreservation methods. Methods Mol Biol. 2013;927:209–15.
Sharma RK, Said T, Agarwal A. Sperm DNA damage and its clinical relevance in assessing reproductive outcome. Asian J Androl. 2004;6:139–48.
Esteves SC, Sharma RK, Thomas Jr AJ, Agarwal A. Effect of swim-up sperm washing and subsequent capacitation on acrosome status and functional membrane integrity of normal sperm. Int J Fertil Womens Med. 2000;45:335–41.
Thomson LK, Fleming SD, Barone K, Zieschang JA, Clark AM. The effect of repeated freezing and thawing on human sperm DNA fragmentation. Fertil Steril. 2010;93:1147–56.
Branco CS, Garcez ME, Pasqualotto FF, Erdtman B, Salvador M. Resveratrol and ascorbic acid prevent DNA damage induced by cryopreservation in human semen. Cryobiology. 2009;60:235–7.
Li Z, Lin Q, Liu R, Xiao W, Liu W. Protective Effects of Ascorbate and Catalase on Human Spermatozoa During Cryopreservation. J Androl. 2009;31:437–44.
WHO. Cambridge; Cambridge University Press; 2010. WHO laboratory manual for the examination and processing of human semen.
Esteves SC, Sharma RK, Thomas Jr AJ, Agarwal A. Improvement in motion characteristics and acrosome status in cryopreserved human spermatozoa by swim-up processing before freezing. Hum Reprod. 2000;15:2173–9.
Counsel M, Bellinge R, Burton P. Vitality of oligozoospermic semen samples is improved by both swim-up and density gradient centrifugation before cryopreservation. J Assist Reprod Genet. 2004;21:137–42.
Abeysundara PK, Dissanayake D, Wijesinghe PS, Perera R, Nishad A. Efficacy of two sperm preparation techniques in reducing non-specific bacterial species from human semen. J Hum Reprod Sci. 2013;6:152–7.
Stanic P, Tandara M, Sonicki Z, Simunic V, Radakovic B, Suchanek E. Comparison of protective media and freezing techniques for cryopreservation of human semen. Eur J Obstet Gynecol Reprod Biol. 2000;91:65–70.
Sion B, Janny L, Boucher D, Grizard G. Annexin V binding to plasma membrane predicts the quality of human cryopreserved spermatozoa. Int J Androl. 2004;27:108–14.
Rodriguez-Martinez H, Larsson B, Pertoft H. Evaluation of sperm damage and techniques for sperm clean-up. Reprod Fertil Dev. 1997;9:297–308.
Muratori M, Porazzi I, Luconi M, Marchiani S, Forti G, Baldi E. AnnexinV binding and merocyanine staining fail to detect human sperm capacitation. J Androl. 2004;25:797–810.
Tavalaee M, Deemeh MR, Arbabian M, Nasr-Esfahani MH. Density gradient centrifugation before or after magnetic-activated cell sorting: which technique is more useful for clinical sperm selection? J Assist Reprod Genet. 2012;29:31–8.
Grunewald S, Paasch U. Sperm selection for ICSI using annexin V. Methods Mol Biol. 2013;927:257–62.
Martin G, Sabido O, Durand P, Levy R. Phosphatidylserine externalization in human sperm induced by calcium ionophore A23187: relationship with apoptosis, membrane scrambling and the acrosome reaction. Hum Reprod. 2005;20:3459–68.
Ricci G, Boscolo R, Martinelli M, Perticarari S. Methods for defecting sperm apoptosis. Fertil Steril. 2009;92(2):e19.
Acknowledgments
The study was financially supported by the Siriraj Grant for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University. I would like to thank Orawan Makemaharn, Suphadtra Phornwilardsiri, Suvana Sangchoo, Sukanya Sriiam, and all of the staff members in the Infertility Unit for their help in conducting the study. Finally, I would like to thank all participants in this research.
Conflict of interests
The study involves no conflict of interests and was financially supported by a Siriraj Grant for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University
Author information
Authors and Affiliations
Corresponding author
Additional information
Author contributions
S.P. contributed to the conception and design of the study, conducted the experiments, performed data analysis and interpretation, and prepared the manuscript. C.N. participated in the experiments and data acquisition, conducted statistical analysis, and was involved in the manuscript preparation. I.T. was involved in revising the manuscript. P.L. and R.S. provided suggestions. All authors read and approved the final manuscript.
Capsule Pre-freezing sperm preparation by swim-up technique improves sperm motility and reduces apoptosis in post-freezing-thawing sperm compared with post-thawing sperm preparation.
Rights and permissions
About this article
Cite this article
Petyim, S., Neungton, C., Thanaboonyawat, I. et al. Sperm preparation before freezing improves sperm motility and reduces apoptosis in post-freezing-thawing sperm compared with post-thawing sperm preparation. J Assist Reprod Genet 31, 1673–1680 (2014). https://doi.org/10.1007/s10815-014-0332-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10815-014-0332-y