Outcomes of intracytoplasmic sperm injection using the zona pellucida-bound sperm or manually selected sperm

  • Rui JinEmail author
  • Junhua BaoEmail author
  • Dawei Tang
  • Fang Liu
  • Guoping Wang
  • Yaqiong Zhao
  • Gang Bai
  • Yongjie Liu
  • Ying Wang
  • Lili Liu
  • Jing Zhao
Assisted Reproduction Technologies



Zona pellucida (ZP)-bound sperm used for intracytoplasmic sperm injection (ICSI) enhances embryo quality, implantation, and clinical pregnancy rates. This study aimed to assess the pregnancy outcomes and clinical significance of ICSI with ZP-bound sperm.


A total of 84 infertile couples who underwent cycles of ICSI following failed in vitro fertilization between June 2012 and February 2014 were enrolled and randomized (1:1): in the treatment group, ICSI was performed using ZP-bound sperm; in the control group, ICSI was performed in a standard manner. Rates of fertilization, cleavage, high-quality embryos, and clinical pregnancy were compared between the two groups.


There were no significant differences in age, infertile period, gonadotrophin dose, number of metaphase II oocytes, and number of embryo transfers between the two groups (P > 0.05). The clinical pregnancy rate was higher in the treatment group than in the control group, but without statistical significance (60.5 vs. 47.6 %, P > 0.05). No significant differences in the rates of fertilization and cleavage were observed (83.0 vs. 81.6 %, and 96.3 vs. 96.5 %, both P > 0.05), but higher rates of high-quality embryos and useable embryos were observed with ZP-bound sperm compared with controls (66.1 vs. 50.8 % and 76.0 vs. 66.3 %, both P < 0.05).


ICSI using ZP-bound sperm might increase the embryo quality and number of useable embryos, possibly improving the clinical pregnancy outcome of ICSI.


In vitro fertilization Intracytoplasmic sperm injection Sperm-zona pellucida binding test Embryo transfer Clinical pregnancy rate Fertilization rate Cleavage rate High-quality embryo rate 



We are grateful to all the participating patients of this study. We thank the staff members of this trial, our colleagues, and all the study staff for their enormous efforts in collecting and ensuring the accuracy and completeness of all the data.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethical Committees of Maternal and Children Health Care Hospital of Yinchuan and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

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


  1. 1.
    Palermo G, Joris H, Devroey P, Van Steirteghem AC. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet. 1992;340(8810):17–8.CrossRefPubMedGoogle Scholar
  2. 2.
    Lanzendorf SE, Maloney MK, Veeck LL, Slusser J, Hodgen GD, Rosenwaks Z. A preclinical evaluation of pronuclear formation by microinjection of human spermatozoa into human oocytes. Fertil Steril. 1988;49(5):835–42.CrossRefPubMedGoogle Scholar
  3. 3.
    Nyboe Andersen A, Carlsen E, Loft A. Trends in the use of intracytoplasmatic sperm injection marked variability between countries. Hum Reprod Update. 2008;14(6):593–604. doi: 10.1093/humupd/dmn032.CrossRefPubMedGoogle Scholar
  4. 4.
    Wen-hao Tang Y-JC. The efficacy and safety of ICS treatment for male infertility. Int J Med Sci - Fam Plann. 2004;23(1):4.Google Scholar
  5. 5.
    Jequier AM. Clinical andrology—still a major problem in the treatment of infertility. Hum Reprod. 2004;19(6):1245–9. doi: 10.1093/humrep/deh269.CrossRefPubMedGoogle Scholar
  6. 6.
    Liu DY. Could using the zona pellucida bound sperm for intracytoplasmic sperm injection (ICSI) enhance the outcome of ICSI? Asian J Androl. 2011;13(2):197–8. doi: 10.1038/aja.2010.179.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Liu F, Qiu Y, Zou Y, Deng ZH, Yang H, de Liu Y. 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(2):815–8. doi: 10.1016/j.fertnstert.2010.09.015.CrossRefPubMedGoogle Scholar
  8. 8.
    Casciani V, Minasi MG, Fabozzi G, Scarselli F, Colasante A, Lobascio AM, et al. Traditional intracytoplasmic sperm injection provides equivalent outcomes compared with human zona pellucida-bound selected sperm injection. Zygote. 2014;22(4):565–70. doi: 10.1017/S0967199413000191.CrossRefPubMedGoogle Scholar
  9. 9.
    Strassburger D, Friedler S, Raziel A, Kasterstein E, Schachter M, Ron-El R. The outcome of ICSI of immature MI oocytes and rescued in vitro matured MII oocytes. Hum Reprod. 2004;19(7):1587–90. doi: 10.1093/humrep/deh236.CrossRefPubMedGoogle Scholar
  10. 10.
    Liu DY, Baker HW. Frequency of defective sperm-zona pellucida interaction in severely teratozoospermic infertile men. Hum Reprod. 2003;18(4):802–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Beaujean N. Epigenetics, embryo quality and developmental potential. Reprod Fertil Dev. 2014;27(1):53–62. doi: 10.1071/RD14309.CrossRefPubMedGoogle Scholar
  12. 12.
    Hammond ER, Stewart B, Peek JC, Shelling AN, Cree LM. Assessing embryo quality by combining non-invasive markers: early time-lapse parameters reflect gene expression in associated cumulus cells. Hum Reprod. 2015. doi: 10.1093/humrep/dev121.Google Scholar
  13. 13.
    O’Neill C, Saunders DM. Assessment of embryo quality. Lancet. 1984;2(8410):1035.CrossRefPubMedGoogle Scholar
  14. 14.
    Ciotti PM, Notarangelo L, Morselli-Labate AM, Felletti V, Porcu E, Venturoli S. First polar body morphology before ICSI is not related to embryo quality or pregnancy rate. Hum Reprod. 2004;19(10):2334–9. doi: 10.1093/humrep/deh433.CrossRefPubMedGoogle Scholar
  15. 15.
    Ola B, Afnan M, Sharif K, Papaioannou S, Hammadieh N, Barratt CL. Should ICSI be the treatment of choice for all cases of in-vitro conception? Considerations of fertilization and embryo development, cost effectiveness and safety. Hum Reprod. 2001;16(12):2485–90.CrossRefPubMedGoogle Scholar
  16. 16.
    Kattera S, Chen C. Short coincubation of gametes in in vitro fertilization improves implantation and pregnancy rates: a prospective, randomized, controlled study. Fertil Steril. 2003;80(4):1017–21.CrossRefPubMedGoogle Scholar
  17. 17.
    Bungum M, Bungum L, Humaidan P. A prospective study, using sibling oocytes, examining the effect of 30 seconds versus 90 minutes gamete co-incubation in IVF. Hum Reprod. 2006;21(2):518–23. doi: 10.1093/humrep/dei350.CrossRefPubMedGoogle Scholar
  18. 18.
    Liu DY, Baker HW. A simple method for assessment of the human acrosome reaction of spermatozoa bound to the zona pellucida: lack of relationship with ionophore A23187-induced acrosome reaction. Hum Reprod. 1996;11(3):551–7.CrossRefPubMedGoogle Scholar
  19. 19.
    Liu DY, Baker HW. Relationship between the zona pellucida (ZP) and ionophore A23187-induced acrosome reaction and the ability of sperm to penetrate the ZP in men with normal sperm-ZP binding. Fertil Steril. 1996;66(2):312–5.CrossRefPubMedGoogle Scholar
  20. 20.
    Martic M, Moses EK, Adams TE, Liu DY, Gook DA, Garrett C, et al. Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line. Asian J Androl. 2004;6(1):3–13.PubMedGoogle Scholar
  21. 21.
    Lin WQ, Lin JJ, Ye BL, Chen YQ, Zhao JZ, Zhou Y, et al. Analysis of pregnancy outcomes after in vitro fertilization-embryo transfer and intracytoplasmic sperm injection. Zhonghua Fu Chan Ke Za Zhi. 2004;39(2):108–11.PubMedGoogle Scholar
  22. 22.
    Papaligoura Z, Panopoulou-Maratou O, Solman M, Arvaniti K, Sarafidou J. Cognitive development of 12 month old Greek infants conceived after ICSI and the effects of the method on their parents. Hum Reprod. 2004;19(6):1488–93. doi: 10.1093/humrep/deh270.CrossRefPubMedGoogle Scholar
  23. 23.
    Avendano C, Franchi A, Taylor S, Morshedi M, Bocca S, Oehninger S. Fragmentation of DNA in morphologically normal human spermatozoa. Fertil Steril. 2009;91(4):1077–84. doi: 10.1016/j.fertnstert.2008.01.015.CrossRefPubMedGoogle Scholar
  24. 24.
    Black M, de Liu Y, Bourne H, Baker HW. 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(2):672–4. doi: 10.1016/j.fertnstert.2009.08.063.CrossRefPubMedGoogle Scholar
  25. 25.
    de Paes Almeida Ferreira Braga D, Laconelli Jr A, de Cassia Savio Figueira R, Madaschi C, Semiao-Francisco L, Borges Jr E. Outcome of ICSI using zona pellucida-bound spermatozoa and conventionally selected spermatozoa. Reprod Biomed Online. 2009;19(6):802–7.CrossRefGoogle Scholar
  26. 26.
    Parmegiani L, Cognigni GE, Bernardi S, Troilo E, Taraborrelli S, Arnone A, et al. Comparison of two ready-to-use systems designed for sperm-hyaluronic acid binding selection before intracytoplasmic sperm injection: PICSI vs. sperm slow: a prospective, randomized trial. Fertil Steril. 2012;98(3):632–7. doi: 10.1016/j.fertnstert.2012.05.043.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Rui Jin
    • 1
    Email author
  • Junhua Bao
    • 1
    Email author
  • Dawei Tang
    • 1
  • Fang Liu
    • 1
  • Guoping Wang
    • 1
  • Yaqiong Zhao
    • 1
  • Gang Bai
    • 1
  • Yongjie Liu
    • 1
  • Ying Wang
    • 1
  • Lili Liu
    • 1
  • Jing Zhao
    • 1
  1. 1.Centre of Assisted ReproductionMaternal and Children Health Care Hospital of YinchuanYinchuanChina

Personalised recommendations