Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
권호 표지
DOI QR코드

DOI QR Code

Evaluation of normal morphology, DNA fragmentation, and hyaluronic acid binding ability of human spermatozoa after using four different commercial media for density gradient centrifugation

  • Lee, Dayong (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital) ;
  • Jee, Byung Chul (Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital)
  • Received : 2018.10.17
  • Accepted : 2018.11.22
  • Published : 2019.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability. Methods: Thirty men with a total motile spermatozoa count > 80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit. Results: The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p< 0.05 for each when compared with 6.9% in raw semen). The DNA fragmentation level was significantly reduced after DGC using PureSperm, PureCeption, and SpermGrad media (6.0%, 6.5%, and 4.9%, respectively; p< 0.05 for each when compared with 11.2% in raw semen), but not after DGC using Sidney media (8.5%, p> 0.05). HA binding ability did not change after DGC using any of the four media. Conclusion: The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.

Keywords

References

  1. Beydola T, Sharma RK, Lee W, Agarwal A. Sperm preparation and selection techniques. In: Rizk B, Aziz N, Agarwal A, Sabanegh E, editors. Male infertility practice. New Delhi: Jaypee Brothers Medical Publishers; 2013. p. 244-51.
  2. Moohan JM, Lindsay KS. Spermatozoa selected by a discontinuous Percoll density gradient exhibit better motion characteristics, more hyperactivation, and longer survival than direct swimup. Fertil Steril 1995;64:160-5. https://doi.org/10.1016/S0015-0282(16)57673-0
  3. Arcidiacono A, Walt H, Campana A, Balerna M. The use of Percoll gradients for the preparation of subpopulations of human spermatozoa. Int J Androl 1983;6:433-45. https://doi.org/10.1111/j.1365-2605.1983.tb00558.x
  4. Iwasaki A, Gagnon C. Formation of reactive oxygen species in spermatozoa of infertile patients. Fertil Steril 1992;57:409-16. https://doi.org/10.1016/S0015-0282(16)54855-9
  5. Mousset-Simeon N, Rives N, Masse L, Chevallier F, Mace B. Comparison of six density gradient media for selection of cryopreserved donor spermatozoa. J Androl 2004;25:881-4. https://doi.org/10.1002/j.1939-4640.2004.tb03157.x
  6. Claassens OE, Menkveld R, Harrison KL. Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation. Hum Reprod 1998;13:3139-43. https://doi.org/10.1093/humrep/13.11.3139
  7. Chiamchanya C, Kaewnoonual N, Visutakul P, Manochantr S, Chaiya J. Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters. Asian J Androl 2010;12:271-7. https://doi.org/10.1038/aja.2009.60
  8. Malvezzi H, Sharma R, Agarwal A, Abuzenadah AM, Abu-Elmagd M. Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial. Reprod Biol Endocrinol 2014;12:121. https://doi.org/10.1186/1477-7827-12-121
  9. Enciso M, Iglesias M, Galan I, Sarasa J, Gosalvez A, Gosalvez J. The ability of sperm selection techniques to remove single- or double-strand DNA damage. Asian J Androl 2011;13:764-8. https://doi.org/10.1038/aja.2011.46
  10. Kim SW, Jee BC, Kim SK, Kim SH. Sperm DNA fragmentation and sex chromosome aneuploidy after swim-up versus density gradient centrifugation. Clin Exp Reprod Med 2017;44:201-6. https://doi.org/10.5653/cerm.2017.44.4.201
  11. Xue X, Wang WS, Shi JZ, Zhang SL, Zhao WQ, Shi WH, et al. Efficacy of swim-up versus density gradient centrifugation in improving sperm deformity rate and DNA fragmentation index in semen samples from teratozoospermic patients. J Assist Reprod Genet 2014;31:1161-6. https://doi.org/10.1007/s10815-014-0287-z
  12. Agarwal A, Majzoub A, Esteves SC, Ko E, Ramasamy R, Zini A. Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios. Transl Androl Urol 2016;5:935-50. https://doi.org/10.21037/tau.2016.10.03
  13. Simon L, Zini A, Dyachenko A, Ciampi A, Carrell DT. A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and intracytoplasmic sperm injection outcome. Asian J Androl 2017;19:80-90.
  14. Zhang LH, Qiu Y, Wang KH, Wang Q, Tao G, Wang LG. Measurement of sperm DNA fragmentation using bright-field microscopy: comparison between sperm chromatin dispersion test and terminal uridine nick-end labeling assay. Fertil Steril 2010;94:1027-32. https://doi.org/10.1016/j.fertnstert.2009.04.034
  15. Worrilow KC, Eid S, Woodhouse D, Perloe M, Smith S, Witmyer J, et al. Use of hyaluronan in the selection of sperm for intracytoplasmic sperm injection (ICSI): significant improvement in clinical outcomes: multicenter, double-blinded and randomized controlled trial. Hum Reprod 2013;28:306-14. https://doi.org/10.1093/humrep/des417
  16. Rashki Ghaleno L, Rezazadeh Valojerdi M, Chehrazi M, Sahraneshin Samani F, Salman Yazdi R. Hyaluronic acid binding assay is highly sensitive to select human spermatozoa with good progressive motility, morphology, and nuclear maturity. Gynecol Obstet Invest 2016;81:244-50. https://doi.org/10.1159/000439530
  17. Johnson SL, Dunleavy J, Gemmell NJ, Nakagawa S. Consistent age-dependent declines in human semen quality: a systematic review and meta-analysis. Ageing Res Rev 2015;19:22-33. https://doi.org/10.1016/j.arr.2014.10.007
  18. Aitken RJ, De Iuliis GN, McLachlan RI. Biological and clinical significance of DNA damage in the male germ line. Int J Androl 2009;32:46-56. https://doi.org/10.1111/j.1365-2605.2008.00943.x
  19. Aitken RJ, De Iuliis GN, Finnie JM, Hedges A, McLachlan RI. Analysis of the relationships between oxidative stress, DNA damage and sperm vitality in a patient population: development of diagnostic criteria. Hum Reprod 2010;25:2415-26. https://doi.org/10.1093/humrep/deq214
  20. Aitken RJ, Clarkson JS, Fishel S. Generation of reactive oxygen species, lipid peroxidation, and human sperm function. Biol Reprod 1989;41:183-97. https://doi.org/10.1095/biolreprod41.1.183

Cited by

  1. Effects of the normal sperm morphology rate on the clinical and neonatal outcomes of conventional IVF cycles vol.52, pp.5, 2019, https://doi.org/10.1111/and.13568
  2. The Author Reply: Effect of Sperm DNA Fragmentation on Embryo Quality in Normal Responder Women in IVF and ICSI vol.61, pp.11, 2020, https://doi.org/10.3349/ymj.2020.61.11.988
  3. Sperm Selection for ICSI: Do We Have a Winner? vol.10, pp.12, 2021, https://doi.org/10.3390/cells10123566