Skip to main content
SpringerLink
Log in

Comparison of intracytoplasmic sperm injection outcome with fresh versus frozen-thawed testicular sperm in men with nonobstructive azoospermia: a systematic review and meta-analysis

  • Assisted Reproduction Technologies
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The purpose of the study is to explore testicular sperm cryopreservation in patients with nonobstructive azoospermia (NOA) whether affect the outcome of subsequent intracytoplasmic sperm injection (ICSI).

Methods

A systematic review and meta-analysis was conducted by searching the MEDLINE and EMBASE databases for relevant published studies in English language (1997–2017). Studies were eligible if they included the comparison of using fresh and frozen-thawed testicular sperm followed by ICSI. Two reviewers independently performed data extraction, quality assessment and assessed the risk of bias. The overall summary risk estimated the number of events. A meta-analysis was conducted using a random effects or fixed effects model analysis according to the test of heterogeneity.

Results

A total of 17 studies with 1,261 ICSI cycles were identified. Analysis of the present data showed no difference in the fertilization outcome when comparing fresh versus frozen-thawed spermatozoa (RR = 1.02, 95% CI 0.86–1.09). Similarly, no difference in CR (RR = 1.01, 95% CI 0.96–1.05), good embryo rate (RR = 1.01, 95% CI 0.95–1.09), and IR (RR = 0.93, 95% CI 0.66–1.30) was observed if the spermatozoa was fresh or frozen-thawed. Finally, no difference in CPR or LBR was noted when using fresh or frozen-thawed cycles were analyzed separately (RR = 1.03, 95% CI 0.86–1.24; RR 1.11, 95% CI 0.88–1.41, respectively).

Conclusions

In men with NOA, the ICSI outcome is not affected by whether the retrieved testicular sperm is fresh or frozen. Sperm cryopreservation ought to be considered in every surgical sperm retrieval case, which remain feasible even in patients with few testicular sperm retrieved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Willott GM. Frequency of azoospermia. Forensic Sci Int. 1982;20:9–10.

    Article  PubMed  CAS  Google Scholar 

  2. Franco G, Misuraca L, Ciletti M, Leonardo C, De Nunzio C, Palminteri E, et al. Surgery of male infertility: an update. RU. 2014;81:154–64.

    Article  Google Scholar 

  3. Ishikawa T. Surgical recovery of sperm in non-obstructive azoospermia. Asian J Androl. 2012;14:109–15.

    Article  PubMed  Google Scholar 

  4. Haimov-Kochman R, Lossos F, Nefesh I, Zentner B-S, Moz Y, Prus D, et al. The value of repeat testicular sperm retrieval in azoospermic men. Fertil Steril. 2009;91:1401–3.

    Article  PubMed  Google Scholar 

  5. Gianaroli L. Diagnostic testicular biopsy and cryopreservation of testicular tissue as an alternative to repeated surgical openings in the treatment of azoospermic men. Hum Reprod. 1999;14:1034–8.

    Article  PubMed  CAS  Google Scholar 

  6. Oates RD, Mulhall J, Burgess C, Cunningham D, Carson R. Fertilization and pregnancy using intentionally cryopreserved testicular tissue as the sperm source for intracytoplasmic sperm injection in 10 men with non-obstructive azoospermia. Hum Reprod. 1997;12:734–9.

    Article  PubMed  CAS  Google Scholar 

  7. Park YS, Lee S-H, Lim CK, Cho JW, Yang KM, Seo JT. Effect of testicular spermatozoa on embryo quality and pregnancy in patients with non-obstructive azoospermia. Syss Biol Reprod Med. 2015;61:300–6.

    Article  Google Scholar 

  8. Madureira C, Cunha M, Sousa M, Neto AP, Pinho MJ, Viana P, et al. Treatment by testicular sperm extraction and intracytoplasmic sperm injection of 65 azoospermic patients with non-mosaic Klinefelter syndrome with birth of 17 healthy children. Andrology. 2014;2:623–31.

    Article  PubMed  CAS  Google Scholar 

  9. Egger M, Smith GD, Schneider M, Minder C. Bias in meta-analysis detected by a simple, graphical test. BMJ. 1997;315:629–34.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Ohlander S, Hotaling J, Kirshenbaum E, Niederberger C, Eisenberg ML. Impact of fresh versus cryopreserved testicular sperm upon intracytoplasmic sperm injection pregnancy outcomes in men with azoospermia due to spermatogenic dysfunction: a meta-analysis. Fertil Steril. 2014;101:344–9.

    Article  PubMed  Google Scholar 

  11. Schlegel PN. Testicular sperm extraction: microdissection improves sperm yield with minimal tissue excision. Hum Reprod. 1999;14:131–5.

    Article  PubMed  CAS  Google Scholar 

  12. Flannigan R, Bach PV, Schlegel PN. Microdissection testicular sperm extraction. Trans Androl Urol. 2017;6:745–52.

    Article  Google Scholar 

  13. Bernie AM, Shah K, Halpern JA, Scovell J, Ramasamy R, Robinson B, et al. Outcomes of microdissection testicular sperm extraction in men with nonobstructive azoospermia due to maturation arrest. Fertil Steril. 2015;104:569–573.e1.

    Article  PubMed  Google Scholar 

  14. Gil-Salom M, Romero J, Menguez Y, Rubio C, De los Santos MJ, Remohe J, et al. Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. Hum Reprod. 1996;11:1309–13.

    Article  PubMed  CAS  Google Scholar 

  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 nonobstructive azoospermia—a comparative study. Fertil Steril. 1997;68:892–7.

    Article  PubMed  CAS  Google Scholar 

  16. Bin W, Wong D, Lu S, Dickstein S, Silva M, Gelety TJ. Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients. J Assis Reprod Genet. 2005;22:389–94.

    Article  Google Scholar 

  17. Agca Y, Critser JK. Cryopreservation of spermatozoa in assisted reproduction. Semin Reprod Med. 2002;20:015–24.

    Article  Google Scholar 

  18. Gómez-Torres MJ, Medrano L, Romero A, Fernández-Colom PJ, Aizpurúa J. Effectiveness of human spermatozoa biomarkers as indicators of structural damage during cryopreservation. Cryobiology. 2017;78:90–4.

    Article  PubMed  CAS  Google Scholar 

  19. Verheyen G, Nagy Z, Joris H, Croo ID, Tournaye H, Steirteghem AV. Quality of frozen-thawed testicular sperm and its preclinical use for intracytoplasmic sperm injection into in vitro-matured germinal-vesicle stage oocytes. Fertil Steril. 1997;67:74–80.

    Article  PubMed  CAS  Google Scholar 

  20. Chatterjee S, Gagnon C. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev. 2001;59:451–8.

    Article  PubMed  CAS  Google Scholar 

  21. Ben-Yosef D, Yogev L, Hauser R, Yavetz H, Azem F, Yovel I, et al. Testicular sperm retrieval and cryopreservation prior to initiating ovarian stimulation as the first line approach in patients with non-obstructive azoospermia. Hum Reprod. 1999;14:1794–801.

    Article  PubMed  CAS  Google Scholar 

  22. Konc J, Kanyó K, Cseh S. The effect of condition/state of testicular spermatozoa injected to the outcome of TESE–ICSI–ET cycles. Eur J Obstet Gynecol Reprod Bio. 2008;141:39–43.

    Article  Google Scholar 

  23. de Oliveira NM. Pregnancy with frozen-thawed and fresh testicular biopsy after motile and immotile sperm microinjection, using the mechanical touch technique to assess viability. Hum Reprod. 2004;19:262–5.

    Article  PubMed  Google Scholar 

  24. Nijs M, Vanderzwalmen P, Vandamme B, Segal-Bertin G, Lejeune B, Segal L, et al. Andrology: fertilizing ability of immotile spermatozoa after intracytoplasmic sperm injection. Hum Reprod. 1996;11:2180–5.

    Article  PubMed  CAS  Google Scholar 

  25. Hauser R. Severe hypospermatogenesis in cases of nonobstructive azoospermia: should we use fresh or frozen testicular spermatozoa? J Androl. 2005;26:772–8.

    Article  PubMed  Google Scholar 

  26. Miah A, Salma U, Schellander K. Roles of relaxin on in vitro sperm physiology. Reprod Domest Anim. 2015;50:125–5.

  27. Aitken RJ, Mattei A, Irvine S. Paradoxical stimulation of human sperm motility by 2-deoxyadenosine. Reproduction. 1986;78:515–27.

    Article  CAS  Google Scholar 

  28. Stephens TD, Brooks RM, Carrington JL, Cheng L, Carrington AC, Porr CA, et al. Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. J Equine Vet Sci. 2013;33:615–21.

    Article  Google Scholar 

  29. Stanic P, Sonicki Z, Suchanek E. Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa. Int J Androl. 2002;25:186–90.

    Article  PubMed  CAS  Google Scholar 

  30. Ebner T, Tews G, Mayer RB, Ziehr S, Arzt W, Costamoling W, et al. Pharmacological stimulation of sperm motility in frozen and thawed testicular sperm using the dimethylxanthine theophylline. Fertil Steril. 2011;96(6):1331–6.

    Article  PubMed  CAS  Google Scholar 

  31. Simopoulou M, Gkoles L, Bakas P, Giannelou P, Kalampokas T, Pantos K, et al. Improving ICSI: a review from the spermatozoon perspective. Syst Bio Reprod Med. 2016;62:359–71.

    Article  Google Scholar 

  32. Kovacic B. Clinical use of pentoxifylline for activation of immotile testicular sperm before ICSI in patients with azoospermia. J Androl. 2006;27:45–52.

    Article  PubMed  CAS  Google Scholar 

  33. Rossato M, Virgilio FD, Foresta C. Involvement of osmo-sensitive calcium influx in human sperm activation. Mol Hum Reprod. 1996;2:903–9.

    Article  PubMed  CAS  Google Scholar 

  34. Stanger JD, Vo L, Yovich JL, Almahbobi G. Hypo-osmotic swelling test identifies individual spermatozoa with minimal DNA fragmentation. Reprod BioMed Online. 2010;21:474–84.

    Article  PubMed  Google Scholar 

  35. Agameya AF, Sallam HN. The value of sperm velocity, strict morphology and the hypo-osmotic swelling test as predictors of the sperm fertilization potential. Fertil Steril. 2014;101:e8.

    Google Scholar 

  36. Sallam HN, Farrag A, Agameya A-F, El-Garem Y, Ezzeldin F. The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study. Hum Reprod. 2005;20:3435–40.

    Article  PubMed  Google Scholar 

  37. Check JH, Tubman A, Wilson C. Intracytoplasmic sperm injection allows normal pregnancy rates for males ≥ 40 with low hypo-osmotic swelling test scores even when complicated by very low motility percentage. Fertil Steril. 2011;95:S28.

    Article  Google Scholar 

  38. Hossain A, Osuamkpe C, Hossain S, Phelps JY. Spontaneously developed tail swellings (SDTS) influence the accuracy of the hypo-osmotic swelling test (HOS-test) in determining membrane integrity and viability of human spermatozoa. J Assis Reprod Genet. 2009;27:83–6.

    Article  Google Scholar 

  39. Soares JB, Glina S, Antunes N, Wonchockier R, Galuppo AG, Mizrahi FE. Sperm tail flexibility test: away to select viable immobile spermatozoa for ICSI. J Urol. 2004;171:363–4.

    Article  Google Scholar 

  40. Nordhoff V. How to select immotile but viable spermatozoa on the day of intracytoplasmic sperm injection? An embryologist’s view. Andrology. 2014;3:156–62.

    Article  PubMed  Google Scholar 

  41. Aktan TM, Montag M, Duman S, Gorkemli H, Rink K, Yurdakul T. Use of a laser to detect viable but immotile spermatozoa. Andrologia. 2004;36:366–9.

    Article  PubMed  CAS  Google Scholar 

  42. Chen H, Feng G, Zhang B, Zhou H, Shu J, Gan X. A successful pregnancy using completely immotile but viable frozen-thawed spermatozoa selected by laser. Clin Exp Reprod Med. 2017;44:52–5.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Sousa M, Cremades N, Silva J, Oliveira C, Ferraz L, da Silva JT, et al. Predictive value of testicular histology in secretory azoospermic subgroups and clinical outcome after microinjection of fresh and frozen-thawed sperm and spermatids. Hum Reprod. 2002;17:1800–10.

    Article  PubMed  CAS  Google Scholar 

  44. De Croo I, Van der Elst J, Everaert K, De Sutter P, Dhont M. Fertilization, pregnancy and embryo implantation rates after ICSI in cases of obstructive and non-obstructive azoospermia. Hum Reprod. 2000;15:1383–8.

    Article  PubMed  Google Scholar 

  45. Stein A, Shufaro Y, Hader S, Fisch B, Pinkas H. Successful use of the Cryolock device for cryopreservation of scarce human ejaculate and testicular spermatozoa. Andrology. 2015;3:220–4.

    Article  PubMed  CAS  Google Scholar 

  46. Coetzee K, Ozgur K, Berkkanoglu M, Bulut H, Isikli A. Reliable single sperm cryopreservation in cell sleepers for azoospermia management. Andrologia. 2016;48:203–10.

    Article  PubMed  CAS  Google Scholar 

  47. Bocca S, Moussavi V, Brugh V, Morshedi M, Stadtmauer L, Oehninger S. ICSI outcomes in men undergoing TESE for azoospermia and impact of maternal age. Andrologia. 2017;49:e12617.

    Article  Google Scholar 

  48. Mazzilli R, Cimadomo D, Vaiarelli A, Capalbo A, Dovere L, Alviggi E, et al. Effect of the male factor on the clinical outcome of intracytoplasmic sperm injection combined with preimplantation aneuploidy testing: observational longitudinal cohort study of 1,219 consecutive cycles. Fertil Steril. 2017;108:961–72.

    Article  PubMed  Google Scholar 

  49. Vernaeve V, Tournaye H, Osmanagaoglu K, Verheyen G, Van Steirteghem A, et al. Intracytoplasmic sperm injection with testicular spermatozoa is less successful in men with nonobstructive azoospermia than in men with obstructive azoospermia. Fertil Steril. 2003;79:529–33.

    Article  PubMed  Google Scholar 

  50. Martin RH, Greene C, Rademaker AW, Ko E, Chernos J. Analysis of aneuploidy in spermatozoa from testicular biopsies from men with nonobstructive azoospermia. J Androl. 2013;24:100–3.

    Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Nation Natural Science Foundation of China (No. 81501246, No. 81501020) and Graduates'’ Innovation Fund, Huazhong University of Science and Technology (No. 5003540054).

Authors’ roles

ZY made substantial contributions to define the research question, acquisition and analyses of data as well as manuscript preparation. ZY and ZWW designed the strategy for literature search, acquisition and interpretation of data. JY and TW assessed eligibility of studies for inclusion to the systematic review. Statistical analyses were performed by HYJ, HL and ZT. SGW and JHL contributed to revision of the manuscript and final approval. All authors read and approved the final version of the manuscript.

Author information

Authors and Affiliations

  1. Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China

    Zhe Yu, Zhewen Wei, Jun Yang, Tao Wang, Hongyang Jiang, Hao Li, Zhe Tang, Shaogang Wang & Jihong Liu

  2. Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Zhe Yu, Zhewen Wei, Jun Yang, Tao Wang, Hongyang Jiang, Hao Li, Zhe Tang, Shaogang Wang & Jihong Liu

Authors
  1. Zhe Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhewen Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hongyang Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zhe Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shaogang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jihong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jun Yang or Jihong Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yu, Z., Wei, Z., Yang, J. et al. Comparison of intracytoplasmic sperm injection outcome with fresh versus frozen-thawed testicular sperm in men with nonobstructive azoospermia: a systematic review and meta-analysis. J Assist Reprod Genet 35, 1247–1257 (2018). https://doi.org/10.1007/s10815-018-1206-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-018-1206-5

Keywords

Search

Navigation