Skip to main content
SpringerLink
Log in

Fertilization with human sperm bound to zona pellucida by pressing onto the oocyte membrane

  • Research Article
  • Published:
Human Cell Aims and scope Submit manuscript

Abstract

This study aimed to determine whether fertilization can be obtained by assisted fusion of oocyte and sperm without breaking the oocyte membrane. A total of 79 infertile couples, each with at least one unfertilized oocyte after in vitro fertilization (IVF), were recruited. Sperm collected from the zona pellucida (ZP) were pressed onto the membrane of unfertilized oocytes at either 6 h or 24 h after IVF, a procedure that we designated as assisted sperm fusion insemination (ASFI). The results of ASFI were compared with those obtained in a previous trial on oocytes in which rescue intracytoplasmic sperm injection (ICSI) was performed at 6 h after IVF. Acrosome reaction (AR) rate of sperm bound to ZP, fertilization rate, degeneration rate, and blastocyst formation rate were evaluated. The AR rate of sperm collected from the ZP was significantly higher than that of the motile sperm recovered from around the oocytes but not bound to the ZP after IVF (98.0% vs. 28.6%). ASFI which was performed at 6 h after IVF yielded a mean fertilization rate of 73.4% (58/79), a degeneration rate of 0% (0/79) and a blastocyst formation rate of 60.8% (31/51). Rescue ICSI which was performed at 6 h after IVF yielded a mean fertilization rate of 70.0% (70/100), a degeneration rate of 4% (4/100) and a blastocyst formation rate of 42.4% (25/59). Binding of sperm to the ZP typically results in AR. ASFI with acrosome-reacted sperm collected from the ZP yielded the fertilization rates similar to those obtained with rescue ICSI.

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

Similar content being viewed by others

References

  1. Nagy ZP, Joris H, Liu J, Staessen C, Devroey P, van Steirteghem AC. Fertilization and early embryology: intracytoplasmic single sperm injection of 1-day-old unfertilized human oocytes. Hum Reprod. 1993;8:2180–4.

    Article  CAS  Google Scholar 

  2. Park KS, Song HB, Chun SS. Late fertilization of unfertilized human oocytes in in vitro fertilization and intracytoplasmic sperm injection cycles: conventional insemination versus ICSI. J Assist Reprod Genet. 2000;17:419–24.

    Article  CAS  Google Scholar 

  3. Lundin K, Sjögren A, Hamberger L. Reinsemination of one-day-old oocytes by use of intracytoplasmic sperm injection. Fertil Steril. 2016;66:118–21.

    Article  Google Scholar 

  4. Goudakou M, Kalogeraki A, Matalliotakis I, Panagiotidis Y, Gullo G, Prapas Y. Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles. Reprod Biomed Online. 2012;24:148–52.

    Article  Google Scholar 

  5. Nagy ZP, Staessen C, Liu J, Joris H, Devroey P, Van Steirteghem AC. Prospective, auto-controlled study on reinsemination of failed-fertilized oocytes by intracytoplasmic sperm injection. Fertil Steril. 1995;64:1130–5.

    Article  CAS  Google Scholar 

  6. Chen C, Kattera S. Rescue ICSI of oocytes that failed to extrude the second polar body 6 h post-insemination in conventional IVF. Hum Reprod. 2003;18:2118–211.

    Article  Google Scholar 

  7. Ohgi S, Hagihara C, Anakubo H, Yanaihara A. A comparison of the clinical outcomes of embryos derived from intracytoplasmic sperm injection after early fertilization check and conventional insemination using sibling oocytes. Arch Gynecol Obstet. 2016;293:887–92.

    Article  Google Scholar 

  8. Laws-King A, Trounson A, Sathananthan H, Kola I. Fertilization of human oocytes by microinjection of a single spermatozoon under the zona pellucida. Fertil Steril. 1987;48:637–42.

    Article  CAS  Google Scholar 

  9. Palermo G, Joris H, Derde M-P, Camus M, Devroey P, Van Steirteghem A. Sperm characteristics and outcome of human assisted fertilization by subzonal insemination and intracytoplasmic sperm injection. Fertil Steril. 1993;59:826–35.

    Article  CAS  Google Scholar 

  10. Kobayashi K, Okuyama M, Fujimoto G, Ogawa S, Rothman CM, Hill DL. Subzonal insemination with a single spermatozoon using manipulation assisted sperm adhesion onto the ooplasmic membrane in mouse ova. Mol Reprod Dev. 1992;31:223–9.

    Article  CAS  Google Scholar 

  11. WHO. WHO laboratory manual for the examination and processing of human semen. 5th edn. World Health. 2010. Vol. 5. p. 286.

  12. Aoki Y, Tsujimura A, Nagashima Y, Hiramatsu I, Uesaka Y, Nozaki T, et al. Effect of Lepidiummeyenii on in vitro fertilization via improvement in acrosome reaction and motility of mouse and human sperm. Reprod Med Biol. 2019;18:57–64.

    Article  CAS  Google Scholar 

  13. Veeck LL. Oocyte assessment and biological performance. Ann N Y Acad Sci. 1988;541:259–74.

    Article  CAS  Google Scholar 

  14. Gardner DK, Schoolcraft WB. In vitro culture of human blastocyst. In: Jansen R, Mortimer D, editors. Towards reproductive certainty: infertility and genetics beyond 1999. Carnforth: Parthenon Press; 1999. p. 378–388.

    Google Scholar 

  15. R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. [Internet]. 2018. https://www.r-project.org. Accessed 10 May 2019.

  16. Liu DY, Baker HWG. Acrosome status and morphology of human spermatozoa bound to the zona pellucida and oolemma determined using oocytes that failed to fertilize in vitro. Hum Reprod. 1994;9:673–9.

    Article  CAS  Google Scholar 

  17. Francavilla F, Marrone V, Romano R, Properzi G, Santucci R, Ruvolo G. Interference of antisperm antibodies with the induction of the acrosome reaction by zona pellucida (ZP) and its relationship with the inhibition of ZP binding. Fertil Steril. 1997;67:1128–33.

    Article  CAS  Google Scholar 

  18. Liu DY, Garrett C, Baker HWG. Low proportions of sperm can bind to the zona pellucida of human oocytes. Hum Reprod. 2003;18:2382–9.

    Article  CAS  Google Scholar 

  19. Yanagida K. Issues of ICSI as a procedure. J Mamm Ova Res. 2013;30:127–34.

    Article  Google Scholar 

  20. De Almeida Ferreira Braga DP, Iaconelli A, De Cássia Sávio Figueira R, Madaschi C, Semiaão-Francisco L, Borges E. Outcome of ICSI using zona pellucida-bound spermatozoa and conventionally selected spermatozoa. Reprod Biomed Online. 2009;19:802–7.

    Article  Google Scholar 

  21. Black M, Liu DY, Bourne H, Baker HWG. Comparison of outcomes of conventional intracytoplasmic sperm injection and intracytoplasmic sperm injection using sperm bound to the zona pellucida of immature oocytes. Fertil Steril. 2010;93:672–4.

    Article  Google Scholar 

  22. Liu F, Qiu Y, Zou Y, Deng ZH, Yang H, Liu DY. Use of zona pellucida-bound sperm for intracytoplasmic sperm injection produces higher embryo quality and implantation than conventional intracytoplasmic sperm injection. Fertil Steril. 2011;95:815–8.

    Article  Google Scholar 

  23. Patrat C, Serres C, Jouannet P. The acrosome reaction in human spermatozoa. Biol cell. 2000;92:255–66.

    Article  CAS  Google Scholar 

  24. Harper CV, Barratt CLR, Publicover SJ, Kirkman-Brown JC. Kinetics of the progesterone-induced acrosome reaction and its relation to intracellular calcium responses in individual human spermatozoa. Biol Reprod. 2006;75:933–9.

    Article  CAS  Google Scholar 

  25. Harper CV, Cummerson JA, White MRH, Publicover SJ, Johnson PM. Dynamic resolution of acrosomal exocytosis in human sperm. J Cell Sci. 2008;121:2130–5.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Graduate School of Health Sciences, Gunma Paz University, 1-7-1 Tonyamachi, Takasaki, Gunma, 370-0006, Japan

    Shota Hatakeyama, Yasuyuki Araki & Yasuhisa Araki

  2. Yanaihara Women’s Clinic, 1-26-29 Ofuna, Kamakura, Kanagawa, 247-0056, Japan

    Shota Hatakeyama, Shirei Ohgi & Atsushi Yanaihara

Authors
  1. Shota Hatakeyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yasuyuki Araki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shirei Ohgi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Atsushi Yanaihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yasuhisa Araki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shota Hatakeyama.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the national committee, and were approved by the Ethics Committee at the Yanaihara Women’s Clinic (YW18-02) and Gunma Paz University (PAZ18-22).

Informed consent

Informed consent was obtained from all individual participants included in the study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hatakeyama, S., Araki, Y., Ohgi, S. et al. Fertilization with human sperm bound to zona pellucida by pressing onto the oocyte membrane. Human Cell 33, 521–527 (2020). https://doi.org/10.1007/s13577-020-00348-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13577-020-00348-4

Keywords

Search

Navigation