Investigation of Time-Dependent Changes in Spermatozoa DNA Condensation and Progressive Motility

Emine Aksoy 1, Müşerref Sultan Mermer 1, Duygu Dursunoğlu 2 * , Fatih Kara 3
More Detail
1 Konya Hospital of Education and Research, IVF Center, Konya
2 Selcuk University, Medicine Faculty, Department of Histology-Embryology, Konya
3 Selcuk University, Medicine Faculty, Department of Public Health, Konya
* Corresponding Author
EUR J BASIC MED SCI, Volume 3, Issue 4, pp. 70-77.
Download Full Text (PDF)


This study aimed to investigate time-dependent changes in spermatozoa DNA condensation and progressive motility and the correlation between DNA condensation and progressive motility of spermatozoa. Semen samples collected from 30 male patients were subdivided into two aliquots. The first aliquot was left unprocessed and labelled ‘raw’. The second aliquot was processed by a swim-up method and was evaluated at the 0th, 3rd and 24th hours. All samples were scored for routine semen analysis. Spermatozoa DNA condensation was evaluated by using the acridine orange and diff-quick stains. The percentages of the changes in the progressive motility and DNA condensation of spermatozoa were estimated. There were no significant differences between samples of the raw and the 0th, 3rd and 24th hours after processing in the percentage of spermatozoa DNA condensation. In all patients, the percentage of spermatozoa progressive motility was higher at the 0th and 3rd hours after processing than in the raw samples. There was no correlation between the percentages of the changes in spermatozoa DNA condensation and progressive motility. Spermatozoa selection can be performed reliably at the 3rd hour after processing of spermatozoa for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) applications. In addition, it is possible to say that the obtaining and processing of spermatozoa that can be performed on the day before the ovum pick-up (OPU) when it is required would not cause to any change in spermatozoa DNA condensation.


Aksoy E, Mermer MS, Dursunoğlu D, Kara F. Investigation of Time-Dependent Changes in Spermatozoa DNA Condensation and Progressive Motility. Eur J Basic Med Sci. 2013;3(4):70-7.


  • Tomlinson MJ, Kessopoulou E, Barratt CL. The diagnostic and prognostic value of traditional semen parameters. J Androl 1999; 20: 588-93.
  • Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002; 23: 25-43.
  • Smit M, Dohle GR, Hop WC, Wildhagen MF, Weber RF, Romijn JC. Clinical correlates of the biological variation of sperm DNA fragmentation in infertile men attending an andrology outpatient clinic. Int J Androl 2007; 30: 48-55.
  • Sakkas D, Mariethoz E, Manicardi GC, Bizzaro D, Bianchi PG, Bianchi U. Origin of DNA damage in ejaculated human spermatozoa. Rev Reprod 1999; 4: 31-7.
  • Saleh RA, Agarwal A, Nelson DR, Nada EA, El-Tonsy MH, Alvarez JG et al. Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study. Fertil Steril 2002; 78: 313-8.
  • Manicardi GC, Tombacco A, Bizzaro D, Bianchi U, Bianchi PG, Sakkas D. DNA strand breaks in ejaculated human spermatozoa: comparison of susceptibility to the nick translation and terminal transferase assays. Histochem J 1998; 30: 33-9.
  • Gorczyca W, Traganos F, Jesionowska H, Darzynkiewicz Z. Presence of strand breaks and increased sensitivity of DNA in situ to denaturation in abnormal human sperm cells: analogy to apoptosis of somatic cells. Exp Cell Res 1993; 207: 202-5.
  • Sakkas D, Urner F, Bizzaro D, Bianchi PG, Wagner I, Jacquenoud N et al. Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection (ICSI). Hum Reprod 1996; 11: 837-43.
  • Bianchi PG, Manicardi GC, Bizzaro D, Campana A, Bianchi U, Sakkas D. The use of the GC specific fluorochrome chromomycin A3 (CMA3) as an indicator of poor sperm quality. J Assist Reprod Genet 1996; 13: 246-50.
  • Moskovtsev SI, Mullen BM. The clinical utility of the evaluation of sperm chromatin. In: Carrell DT, Peterson CM editors. Reproductive endocrinology and infertility: integrating modern clinical and laboratory practice. New York: Springer Publishers; 2010. p.467-83.
  • Roux C, Tripogney C, Joanne C, Bresson JL. Nuclear quality of the spermatozoon: exploration tests of the chromatin of human spermatozoa (nuclear proteins). Gynecol Obstet Fertil 2004; 32: 792-8.
  • Chohan KR, Griffin JT, Laframboise M, De Jonge CJ, Carrell DT. Comparison of chromatin assays for DNA fragmentation evaluation in human sperm. J Androl 2006; 27: 53-9.
  • World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. Cambridge: Cambridge University Press; 1999.
  • Spano M, Bonde JP, Hjollund HI, Kolstad HA, Cordelli E, Leter G. Sperm chromatin damage impairs human fertility. The Danish first pregnancy planner study team. Fertil Steril 2000; 73: 43-50.
  • Sharma RK, Said T, Agarwal A. Sperm DNA damage and its clinical relevance in assessing reproductive outcome. Asian J Androl 2004; 6: 139-48.
  • Bungum M, Humaidan P, Spano M, Jepson K, Bungum L, Giwercman A. The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI. Hum Reprod 2004; 19: 1401-8.
  • Benchaib M, Lornage J, Mazoyer C, Lejeune H, Salle B, François Guerin J. Sperm deoxyribonucleic acid fragmentation as a prognostic indicator of assisted reproductive technology outcome. Fertil Steril 2007; 87: 93-100.
  • Bungum M, Humaidan P, Axmon A, Spano M, Bungum L, Erenpreiss J et al. Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome. Hum Reprod 2007; 22: 174-9.
  • Speyer BE, Pizzey AR, Ranieri M, Joshi R, Delhanty JD, Serhal P. Fall in implantation rates following ICSI with sperm with high DNA fragmentation. Hum Reprod 2010; 25: 1609-18.
  • Gandini L, Lombardo F, Paoli D, Caruso F, Eleuteri P, Leter G et al. Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage. Hum Reprod 2004; 19: 1409-17.
  • Yılmaz S, Demiroglu A, Yılmaz E, Sofuoglu K, Delikara N, Kutlu P. Effects of sperm DNA fragmentation on semen parameters and ICSI outcome determined by an improved SCD test, halosperm. Int J Fertil Steril 2010; 4: 73-8.
  • Larson-Cook KL, Brannian JD, Hansen KA, Kasperson KM, Aamold ET, Evenson DP. Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay. Fertil Steril 2003; 80: 895-902.
  • Hammadeh ME, Zeginiadov T, Rosenbaum P, Georg T, Schmidt W, Strehler E. Predictive value of spermatozoa chromatin condensation (aniline blue staining) in the assessment of male fertility. Arch Androl 2001; 46: 99-104.
  • Apedaile AE, Garrett C, Liu DY, Clarke GN, Johnston SA, Baker HW. Flow cytometry and microscopic acridine orange test: relationship with standard semen analysis. Reprod Biomed Online 2004; 8: 398-407.
  • Moskovtsev SI, Willis J, Mullen JB. Age-related decline in sperm deoxyribonucleic acid integrity in patients evaluated for male infertility. Fertil Steril 2006; 85: 496-9.
  • Cohen-Bacrie P, Belloc S, Ménézo YJ, Clement P, Hamidi J, Benkhalifa M. Correlation between DNA damage and sperm parameters: a prospective study of 1,633 patients. Fertil Steril 2009; 91: 1801-5.
  • Irvine DS, Twigg JP, Gordon EL, Fulton N, Milne PA, Aitken RJ. DNA integrity in human spermatozoa: relationships with semen quality. J Androl 2000; 21: 33-44.
  • Sakkas D, Urner F, Bizzaro D, Manicardi G, Bianchi U, Shoukir Y et al. Sperm nuclear DNA damage and altered chromatin structure: effect on fertilization and embryo development. Hum Reprod 1998; 13: 11-9.
  • Tomlinson MJ, Moffatt O, Manicardi GC, Bizzaro D, Afnan M, Sakkas D. Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: implications for assisted conception. Hum Reprod 2001; 16: 2160-5.
  • Moskovtsev SI, Willis J, White J, Mullen JB. Leukocytospermia: relationship to sperm deoxyribonucleic acid integrity in patients evaluated for male factor infertility. Fertil Steril 2007; 88: 737-40.
  • Moskovtsev SI, Willis J, White J, Mullen JB. Sperm DNA damage: correlation to severity of semen abnormalities. Urology 2009; 74: 789-93.
  • Evenson DP, Jost LK, Marshall D, Zinaman MJ, Clegg E, Purvis K et al. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 1999; 14: 1039-49.
  • Sousa AP, Tavares RS, Velez de la Calle JF, Figueiredo H, Almeida V, Almeida-Santos T et al. Dual use of diff-quiklike stains for the simultaneous evaluation of human sperm morphology and chromatin status. Hum Reprod. 2009; 24: 28-36.
  • Teyada RI, Mitchell JC, Norman A, Marik JJ, Friedman S. A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril 1984; 42: 87-91.
  • Duran EH, Gurgan T, Gunalp S, Enginsu ME, Yarali H, Ayhan A. A logistic regression model including DNA status and morphology of spermatozoa for prediction of fertilization in vitro. Hum Reprod. 1998; 13: 1235-9.
  • Ghumman S, Adiga SK, Upadhya D, Kalthur G, Jayaraman V, Rao SB et al. Combination of swim-up and density gradient separation methods effectively eliminate DNA damaged sperm. J Turkish-German Gynecol Assoc 2011; 12: 148-52.
  • Marchesi DE, Biederman H, Ferrara S, Hershlag A, Feng HL. The effect of semen processing on sperm DNA integrity: comparison of two techniques using the novel toluidine blue assay. Eur J Obstet Gynecol Reprod Biol 2010; 151: 176-80.
  • Levran D, Ginath S, Farhi J, Nahum H, Glezerman M, Weissman A. Timing of testicular sperm retrieval procedures and in vitro fertilization intracytoplasmic sperm injection outcome. Fertil Steril 2001; 76: 380-3.
  • Hu Y, Maxson WS, Hoffman DI, Ory SJ, Licht MR, Eager S. Clinical application of intracytoplasmic sperm injection using in vitro cultured testicular spermatozoa obtained the day before egg retrieval. Fertil Steril 1999; 72: 666-9.
  • Jackson RE, Bormann CL, Hassun PA, Rocha AM, Motta EL, Serafini PC et al. Effects of semen storage and separation techniques on sperm DNA fragmentation Fertil Steril 2010; 94: 2626-30.