Advertisement

Journal of Assisted Reproduction and Genetics

, Volume 25, Issue 8, pp 375–381 | Cite as

Clinical outcome of magnetic activated cell sorting of non-apoptotic spermatozoa before density gradient centrifugation for assisted reproduction

  • Enver Kerem Dirican
  • Osman Denizhan Özgün
  • Süleyman Akarsu
  • Kadir Okhan Akın
  • Özge Ercan
  • Mukaddes Uğurlu
  • Çağrı Çamsarı
  • Oya Kanyılmaz
  • Adnan Kaya
  • Ali Ünsal
ASSISTED REPRODUCTION

Abstract

Purpose

Magnetic activated cell sorting (MACS) eliminates apoptotic spermatozoa based on the presence of externalized phosphatidylserine residues. We evaluated the outcome of male fertility treatment when intracytoplasmic sperm injection (ICSI) into human oocytes was performed with non-apoptotic MACS-selected spermatozoa.

Methods

196 couples were treated by ICSI following spermatozoa preparation by MACS (study group; 122 couples) or density gradient centrifugation (DGC) (control group; 74 couples). Fertilization, cleavage, pregnancy, and implantation rates were analyzed.

Results

The percentage of sperm with normal morphology after MACS selection was improved. Cleavage and pregnancy rates were higher, respectively, in the study group than in control. A slightly higher implantation rate was also observed in the study group.

Conclusions

MACS selection of human spermatozoa increased cleavage and pregnancy rates in oligoasthenozoospermic ART cases. This novel method for selecting non-apoptotic spermatozoa for ICSI is safe and reliable, and may improve the assisted reproduction outcome.

Keywords

Apoptosis Spermatozoa Intracytoplasmic sperm injection Pregnancy rate 

References

  1. 1.
    Seli E, Gardner DK, Schoolcraft WB, Moffatt O, Sakkas D. Extent of nuclear DNA damage in ejaculated spermatozoa impacts on blastocyst development after in vitro fertilization. Fertil Steril. 2004;82:378–83. doi: 10.1016/j.fertnstert.2003.12.039.PubMedCrossRefGoogle Scholar
  2. 2.
    Henkel R, Hajimohammad M, Stalf T, Hoogendijk C, Mehnert C, Menkveld R, et al. Influence of deoxyribonucleic acid damage on fertilization and pregnancy. Fertil Steril. 2004;81:965–72. doi: 10.1016/j.fertnstert.2003.09.044.PubMedCrossRefGoogle Scholar
  3. 3.
    Sun JG, Jurisicova A, Casper RF. Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod. 1997;56:602–7. doi: 10.1095/biolreprod56.3.602.PubMedCrossRefGoogle Scholar
  4. 4.
    Ombelet W, Deblaere K, Bosmans E, Cox A, Jacobs P, Janssen M, et al. Semen quality and intrauterine insemination. RBM Online. 2003;7:485–92.PubMedGoogle Scholar
  5. 5.
    Said TM, Grunewald S, Paasch U, Glander HJ, Baumann T, Kriegel C, et al. Advantage of combining magnetic cell separation with sperm preparation techniques. RBM Online. 2005;10(6):740–6.PubMedGoogle Scholar
  6. 6.
    Flesch FM, Gadella BM. Dynamics of the mammalian sperm plasma membrane in the process of fertilization. Biochim Biophys Acta. 2000;1469:197–235.PubMedGoogle Scholar
  7. 7.
    Glander HJ, Schaller J. Binding of annexin V to plasma membranes of human spermatozoa: a rapid assay for detection of membrane changes after cryostorage. Mol Hum Reprod. 1999;5:109–15. doi: 10.1093/molehr/5.2.109.PubMedCrossRefGoogle Scholar
  8. 8.
    Duru NK, Morshedi MS, Schuffner A, Oehninger S. Cryopreservation-thawing of fractionated human spermatozoa is associated with membrane phosphatidylserine externalization and not DNA fragmentation. J Androl. 2001;22:383–424.Google Scholar
  9. 9.
    Shen HM, Dai J, Chia SE, Lim A, Ong CN. Detection of apoptotic alterations in sperm in subfertile patients and their correlations with sperm quality. Hum Reprod. 2002;17:1266–73. doi: 10.1093/humrep/17.5.1266.PubMedCrossRefGoogle Scholar
  10. 10.
    Grunewald S, Paasch U, Glander HJ. Enrichment of non-apoptotic human spermatozoa after cryopreservation by immunomagnetic cell sorting. Cell Tissue Bank. 2001;2:127–33. doi: 10.1023/A:1020188913551.PubMedCrossRefGoogle Scholar
  11. 11.
    Kang JH, Park J. Cell separation technology. APBN. 2005;9(21):1139–2005.Google Scholar
  12. 12.
    van Heerde WL, de Groot PG, Reutelingsperger CP. The complexity of the phospholipid binding protein annexin V. Thromb Haemost. 1995;73:172–9.PubMedGoogle Scholar
  13. 13.
    Glander HJ, Schaller J. Binding of annexin V to plasma membranes of human spermatozoa: a rapid assay for detection of membrane changes after cryostorage. Mol Hum Reprod. 1999;5:109–15. doi: 10.1093/molehr/5.2.109.PubMedCrossRefGoogle Scholar
  14. 14.
    Said TM, Paasch U, Grunewald S, Rasch M, Glander H, Agarwal A. Sperm recovery evaluation following magnetic cell sorting. Fertil Steril. 2005;84:S207. doi: 10.1016/j.fertnstert.2005.07.524.CrossRefGoogle Scholar
  15. 15.
    Said TM, Agarwal A, Erenpreiss J, Mahfouz R, Giwercman A, Evenson DP. Association of early apoptosis with the nuclear integrity and maturity in different seminal fractions. Fertil Steril. 2006;86(3):S499.Google Scholar
  16. 16.
    World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. Cambridge: Cambridge University; 1999.Google Scholar
  17. 17.
    Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF, Veeck LL, et al. New method of evaluating sperm morphology with predictive value for human in vitro fertilization. Urol. 1987;30(3):248–51. doi: 10.1016/0090-4295(87)90246-9.PubMedCrossRefGoogle Scholar
  18. 18.
    Palermo GD, Schlegel PN, Colombero LT, Zaninovic N, Moy F, Rozenwaks Z. Aggressive sperm immobilization prior to intracytoplasmic sperm injection with immature spermatozoa improves fertilization and pregnancy rates. Hum Reprod. 1996;11:1023–9.PubMedGoogle Scholar
  19. 19.
    Sinha Hikim AP, Swerdloff RS. Hormonal and genetic control of germ cell apoptosis in the testis. Rev Reprod. 1999;4:38–47. doi: 10.1530/ror.0.0040038.PubMedCrossRefGoogle Scholar
  20. 20.
    Taylor SL, Weng SL, Fox P, Duran EH, Morshedi MS, Oehninger S, et al. Somatic cell apoptosis markers and pathways in human ejaculated sperm: potential utility as indicators of sperm quality. Mol Hum Reprod. 2004;10:825–34. doi: 10.1093/molehr/gah099.PubMedCrossRefGoogle Scholar
  21. 21.
    Sakkas D, Mariethoz E, St John JC. Abnormal sperm parameters in humans are indicative of an abortive apoptotic mechanism linked to the Fas-mediated pathway. Exp Cell Res. 1999;251:350–5. doi: 10.1006/excr.1999.4586.PubMedCrossRefGoogle Scholar
  22. 22.
    Sakkas D, Moffatt O, Manicardi GC, Mariethoz E, Tarozzi N, Bizzaro D. Nature of DNA damage in ejaculated human spermatozoa and the possible involvement of apoptosis. Biol Reprod. 2002;66:1061–7. doi: 10.1095/biolreprod66.4.1061.PubMedCrossRefGoogle Scholar
  23. 23.
    Barroso G, Morshedi M, Oehninger S. Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. Hum Reprod. 2000;15:1338–44. doi: 10.1093/humrep/15.6.1338.PubMedCrossRefGoogle Scholar
  24. 24.
    Gorczyca W, Traganos F, Jesionowska H, Darzynkiewicz Z. Presence of DNA 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. doi: 10.1006/excr.1993.1182.PubMedCrossRefGoogle Scholar
  25. 25.
    Kasai T, Ogawa K, Mizuno K, Nagai S, Uchida Y, Ohta S, et al. Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential. Asian J Androl. 2002;4:97–103.PubMedGoogle Scholar
  26. 26.
    Weng SL, Taylor SL, Morshedi M, Schuffner A, Duran EH, Beebe S, et al. Caspase activity and apoptotic markers in ejaculated human sperm. Mol Hum Reprod. 2002;8:984–91. doi: 10.1093/molehr/8.11.984.PubMedCrossRefGoogle Scholar
  27. 27.
    Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger CP. A novel assay for apoptosis: flow cytometric detection of phosphatidylserine expression of early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods. 1995;184:39–51. doi: 10.1016/0022-1759(95)00072-I.PubMedCrossRefGoogle Scholar
  28. 28.
    Oosterhuis GJ, Mulder AB, Kalsbeek-Batenburg E, Lambalk CB, Schoemaker J, Vermes I. Measuring apoptosis in human spermatozoa: a biological assay for semen quality? Fertil Steril. 2000;74:245–50. doi: 10.1016/S0015-0282(00)00623-3.PubMedCrossRefGoogle Scholar
  29. 29.
    van Heerde WL, de Groot PG, Reutelingsperger CP. The complexity of the phospholipid binding protein annexin V. Thromb Haemost. 1995;73:172–9.PubMedGoogle Scholar
  30. 30.
    Donnelly ET, O, Connell M, McClure N, Lewis SE. Differences in nuclear DNA fragmentation and mitochondrial integrity of semen and prepared human spermatozoa. Hum Reprod. 2000;15:1552–61. doi: 10.1093/humrep/15.7.1552.PubMedCrossRefGoogle Scholar
  31. 31.
    Marchetti C, Obert G, Deffosez A, Formstecher P, Marchetti P. Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm. Hum Reprod. 2002;17:1257–65. doi: 10.1093/humrep/17.5.1257.PubMedCrossRefGoogle Scholar
  32. 32.
    Said TM, Paasch U, Grunewald S, Rasch M, Glander H, Agarwal A. Impact of apoptosis and DNA damage on sperm chromatin decondensation following intracytoplasmic injection. Fertil Steril. 2005;84:S137. doi: 10.1016/j.fertnstert.2005.07.335.CrossRefGoogle Scholar
  33. 33.
    Dirican EK, Kalender Y, Uzunhisarcikli M, Ogutcu A. Testicular toxicity of dichlorvos and protective roles of vitamins C and E in male rats. 6th International Congress of Turkish Society of Toxicology Antalya, Turkey, 2006.Google Scholar
  34. 34.
    Uzunhisarcıklı M, Kalender Y, Dirican K, Kalender S, Ögütçü A, Büyükkömürcü F. Acute, subacute and subchronic administration of methyl parathion-induced testicular damage in male rats and the protective role of vitamins C and E. Pestic Biochem Physiol. 2007;87(2):115–22. doi: 10.1016/j.pestbp.2006.06.010.CrossRefGoogle Scholar
  35. 35.
    Crosignani PG, Collins J, Cooke ID, Diczfalusy E, Rubin B. Recommendation of the ESHRE workshop on unexplained infertility. Hum Reprod. 1992;8:977–80.Google Scholar
  36. 36.
    Agarwal A, Said TM, Bedaiwy MA, Banerjee J, Alvarez JG. Oxidative stress in an assisted reproductive techniques setting. Fertil Steril. 2006;86(3):503–12. doi: 10.1016/j.fertnstert.2006.02.088.PubMedCrossRefGoogle Scholar
  37. 37.
    Brugnon F, Verheyen G, Janny L, Pouly J, Liebaers I, Vansteirteghem A. Study of apoptosis by DNA fragmentation and phosphatidylserine externalization, and of meiotic segregation in ejaculated sperm of chromosomal translocation carrier patients. Fertil Steril. 2005;84:S61. doi: 10.1016/j.fertnstert.2005.07.147.CrossRefGoogle Scholar
  38. 38.
    Barroso G, Taylor S, Morshedi M, Manzur F, Gaviño F, Oehninger S. Mitochondrial membrane potential integrity and plasma membrane translocation of phosphatidylserine as early apoptotic markers: a comparison of two different sperm subpopulations. Fertil Steril. 2006;85(1):149–54. doi: 10.1016/j.fertnstert.2005.06.046.PubMedCrossRefGoogle Scholar
  39. 39.
    Aziz N, Said TM, Paasch U, Grunewald S, Sharma RK, Agarwal A. The cause why apoptotic sperm have poor morphology profile as assessed by the sperm deformity index (SDI): A prospective study. Fertil Steril. 2006;86(3):S506. doi: 10.1016/j.fertnstert.2006.07.1400.CrossRefGoogle Scholar
  40. 40.
    Said TM, Aziz N, Paasch U, Grunewald S, Glander H, Agarwal A. Elimination of apoptotic sperm as a measure for enhancing morphological quality as assessed by the sperm deformity (SDI). Fertil Steril. 2005;84:S448–9. doi: 10.1016/j.fertnstert.2005.07.1175.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Enver Kerem Dirican
    • 1
  • Osman Denizhan Özgün
    • 2
  • Süleyman Akarsu
    • 3
  • Kadir Okhan Akın
    • 4
  • Özge Ercan
    • 1
  • Mukaddes Uğurlu
    • 5
  • Çağrı Çamsarı
    • 5
  • Oya Kanyılmaz
    • 6
  • Adnan Kaya
    • 2
  • Ali Ünsal
    • 7
  1. 1.EmbryologyMAYA Center for Assisted Reproductive TechnologiesAnkaraTurkey
  2. 2.Obstetrics and GynecologyMAYA Center for Assisted Reproductive TechnologiesAnkaraTurkey
  3. 3.Obstetrics and GynecologyKeçiören Education and Research HospitalAnkaraTurkey
  4. 4.BiochemistryKeçiören Education and Research HospitalAnkaraTurkey
  5. 5.AndrologyMAYA Center for Assisted Reproductive TechnologiesAnkaraTurkey
  6. 6.Public HealthMAYA Center for Assisted Reproductive TechnologiesAnkaraTurkey
  7. 7.UrologyKeçiören Education and Research HospitalAnkaraTurkey

Personalised recommendations