Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients

  • Bin Wu
  • Diana Wong
  • Suzhen Lu
  • Stephen Dickstein
  • Marta Silva
  • Timothy J. Gelety
Andrology

Abstract

Purpose: To optimize the use of fresh and frozen-thawed testicular biopsy specimens from patients with azoospermia.

Methods: Fifty-one patients suffering from obstructive and non-obstructive azoospermia underwent testicular sperm extraction (TESE). The specimens were divided and used for either in vitro maturation or freezing for a future intracytoplasmic sperm injection (ICSI) cycle.

Results: At initial testicular sperm extraction, very few motile spermatozoa were seen. After 24 h of in vitro maturation, sperm motility increased remarkably, with a maximum motility rate seen between 48 and 72 h of culture. Motile spermatozoa were observed up to 120 h in culture. In the 22 fresh ICSI cycles, a total of 294 oocytes were injected using motile sperm and 212 oocytes demonstrated normal 2PN formation (fertilization rate, 72.1%). In 36 frozen-thawed ICSI cycles, a total of 454 oocytes were injected and 302 oocytes became 2PN (66.5%). On day 3, high quality embryos were observed in 54.2% of fresh cycles and 54.1% of frozen cycles (P > 0.05). The clinical pregnancy rate did not show a significant difference between using fresh (59%) and frozen (55.5%) testicular biopsy sperm for ICSI (P > 0.05), but the embryo implantation rates did differ significantly between fresh (29.5%) and frozen-thawed (22.2%) cycles (P < 0.05). A total of 33 healthy babies have been born from 22 women, giving birth after 58 embryo transfer attempts (38%).

Conclusion: The freezing and in vitro culturing of testicular biopsy tissue is a very reliable approach for the management of testicular biopsy specimens from azoospermic patients, and offers the possibility of several treatments of IVF/ICSI from a single sample.

Keywords

Cryopreservation in vitro maturation testicular sperm extraction 

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References

  1. 1.
    Palermo G, Cohen J, Alikani M, Alder A, Rosenwaks Z: Intracytoplasmic sperm injection: A novel treatment for all forms of male infertility. Fertil Steril 1995;63:1231–1245PubMedGoogle Scholar
  2. 2.
    Romero J, Remohi J, Minguez Y, Rubio C, Pellicer A, Gil-Salom M: Fertilization after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. Fertil Steril 1996;65:877–879PubMedGoogle Scholar
  3. 3.
    Angelopoulos T, Adler A, Krey L, Licciardi F, Noyes N, McCullough A: Enhancement or initiation of testicular sperm motility by in vitro culture of testicular tissue. Fertil Steril 1999;71:240–243CrossRefPubMedGoogle Scholar
  4. 4.
    Emiliani S, Van den Bergh M, Vannin A, Biramane J, Verdoodt M, Englert Y: Increased sperm motility after in vitro culture of testicular biopsies from obstructive azoospermic patients results in better post-thawed recovery rate. Hum Reprod 2000;12:2371–2374Google Scholar
  5. 5.
    Pasqualotto F, Rossi-Ferragut R, Rocha C, Iaconelli A, Borges E: Outcome of in vitro fertilization and intracytoplasmic injection of epididymal and testicular sperm obtained from patients with obstructive and non-obstructive azoospermia. J Urol 2002;167:1753–1756PubMedGoogle Scholar
  6. 6.
    Schoor R, Elhanbly S, Niederberger C, Ross L: The role of testicular biopsy in the modern management of male infertility. J Urol 2002;167:197–200PubMedGoogle Scholar
  7. 7.
    Jow WW, Steckel J, Schelgel PN, Magid MS, Goldstein M: Motile sperm in human testis biopsy specimen. J Androl 1993;14:194–198PubMedGoogle Scholar
  8. 8.
    Lacham-Kaplan O, Trounson A: The effects of the sperm motility activators 2-deoxyadenosine and pentoxifylline used for sperm microinjection on mouse and human embryo development. Hum Reprod 1993;8:945–952PubMedGoogle Scholar
  9. 9.
    Nagy Z, Liu J, Cecile J, Silber S, Devroey P, Van Steirteghem A: Using ejaculated, fresh, and frozen-thawed epididymal and testicular spermatozoa gives rise to comparable results after intracytoplasmic sperm injection. Fertil Steril 1995;63:808–815PubMedGoogle Scholar
  10. 10.
    Liu J, Tsai Y, Katz E: Outcome of in vitro culture of fresh and frozen thawed human testicular spermatozoa. Hum Reprod 1997;12:1667–1672PubMedGoogle Scholar
  11. 11.
    Craft I, Tsirigotis M, Zhu J: In vitro culture of testicular sperm (letter). Lancet 1995;346:1439CrossRefGoogle Scholar
  12. 12.
    Zhu J, Tsirigotis M, Pelelcamos M: In vitro maturation of human testicular spermatozoa. Hum Reprod 1996;11:231–232PubMedGoogle Scholar
  13. 13.
    Liu J, Garcia E, Baramki A: The difference in outcome of in vitro culture of human testicular spermatozoa between obstructive and non-obstructive azoospermia. Hum Reprod 1996;11:1587–1588PubMedGoogle Scholar
  14. 14.
    Allan J, Cotman A: A new method for freezing testicular biopsy sperm: Three pregnancies with sperm extracted from cryopreserved sections of seminiferous tubule. Fertil Steril 1997;68:741–744CrossRefPubMedGoogle Scholar
  15. 15.
    Friedler S, Raziel A, Soffer Y, Strassburger D, Komarovsky D, Ron-El R: Intracytoplasmic injection of fresh and cryopreserved testicular spermatozoa in patients with non-obstructive azoospermia—A comparative study. Fertil Steril 1997;68:892–897CrossRefPubMedGoogle Scholar
  16. 16.
    Grabbe E, Verheyen G, Taureye H, Van Steirteghem A: Freezing of testicular tissue as a minced suspension preserves sperm quality better than whole-biopsy freezing when glycerol is used as cryoprotectant. Int J Androl 1999;22:43–48Google Scholar
  17. 17.
    Park YS, Lee SH, Song SJ, Jun JH, Koong MK, Seo JT: Influence of motility on the outcome of in vitro fertilization/intracytoplasmic sperm injection with fresh vs. frozen testicular sperm from men with obstructive azoospermia. Fertil Steril 2003;80:526–530PubMedGoogle Scholar
  18. 18.
    Aoki VW, Wilcox AL, Thorp C, Hamilton BD, Carrell DT: Improved in vitro fertilization embryo quality and pregnancy rates with intracytoplasmic sperm injection of sperm from fresh testicular biopsy samples vs. frozen biopsy samples. Fertil Steril 2004;82:1532–1535CrossRefPubMedGoogle Scholar
  19. 19.
    Thompson-Cree MEM, McClure N, Donnelly Eilish T, Steele Kristine E, Lewis Sheena EM: The effects of cryopreservation on testicular sperm nuclear DNA fragmentation and its relationship with assisted conception outcome following intracytoplasmic sperm injection. Reprod Biomed Online 2003;7(4):449–455Google Scholar
  20. 20.
    Anderson AR, Wiemer KE, Weikert ML, Kyslinger ML: Fertilization, embryonic development and pregnancy losses with intracytoplasmic sperm injection for surgically-retrieved spermatozoa. Reprod Biomed Online 2002;5(2):142–147Google Scholar
  21. 21.
    Said TM, Agarwal A, Sharma RK, Thomas AJ Jr, Sikka SC: Impact of sperm morphology on DNA damage caused by oxidative stree induced by beta-nicotinamide adenine dinucleotide phosphate. Fertil Steril 2005;83:95–103PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Bin Wu
    • 1
    • 2
  • Diana Wong
    • 1
  • Suzhen Lu
    • 1
  • Stephen Dickstein
    • 1
  • Marta Silva
    • 1
  • Timothy J. Gelety
    • 1
  1. 1.Arizona Center for Reproductive Endocrinology and InfertilityTucson
  2. 2.College of Animal Science and Veterinary MedicineNorthwest A&F UniversityYanglingP.R. China

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