The hemizona assay (HZA): A predictor of human sperm fertilizing potential in in vitro fertilization (IVF) treatment

  • Daniel R. Franken
  • Sergio Oehninger
  • Lani Johnson Burkman
  • Charles C. Coddington
  • Thinus F. Kruger
  • Zev Rosenwaks
  • Anibal A. Acosta
  • Gary D. Hodgen
Special Contributions


The hemizona assay (HZA) was developed to assess human sperm fertilizing potential. This blinded study investigated the relationship between sperm binding to the hemizona and in vitro fertilization (IVF) success (36 patients). Nonliving human oocytes were recovered from excised ovaries and stored. Each zona pellucida was cut into equal hemispheres by micromanipulation. For the HZA, one droplet exposed a hemizona to abnormal spermatozoa, while the control droplet contained the matching hemizona and spermatozoa from normal semen. After 4 hr, the number of tightly bound spermatozoa was counted. Binding to the hemizona was significantly higher for those having IVF success (mean of 36.1±7, versus 10.4±4 from the failure group;P<0.05). Fewer sperm from the failure group had a strictly normal morphology (3,2 versus 12.7%;P<0.05, Kruger method). Tight zona binding was significantly correlated with the percentage motile sperm, percentage normal morphology, and seminal sperm concentration. These results enhanced our confidence that the HZA is diagnostic for identification of patients at high risk of failing to achieve fertilization in vitro.

Key Words

zona pellucida human fertility sperm binding in vitro fertilization (IVF) 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Yanagimachi R: Mechanism of fertilization in mammals.In Fertilization and Embryonic Development in Vitro. L Mastroianni, JB Biggers (eds). New York, Plenum, 1981, pp 81–182Google Scholar
  2. 2.
    Florman HM, Bechtol KB, Wasserman PM: Enzymatic disection of the functions of the mouse egg's receptor for sperm. Dev Biol 1984;106:243–255Google Scholar
  3. 3.
    Gould JE, Overstreet JW, Yanagimachi H, Yanagimachi R, Katz DF, Hanson FW: What functions of the sperm cell are measured by in vitro fertilization of zona-free hamster eggs? Fertil Steril 1983;40:344–352Google Scholar
  4. 4.
    Overstreet JW, Hembree WC: Penetration of zona pellucida of nonliving human oocytes by human spermatozoa in vitro. Fertil Steril 1976;27:815–831Google Scholar
  5. 5.
    Overstreet JW, Yanagimachi R, Katz DF, Havashi K, Hanson FW: Penetration of human spermatozoa into the human zona pellucida and the zona-free hamster egg: A study of fertile donors and infertile patients. Fertil Steril 1980;33:534–542Google Scholar
  6. 6.
    Singer SL, Lambert H, Overstreet JW, Hanson FW, Yanagimachi R: The kinetics of human sperm binding to the human zona pellucida and zona-free hamster oocyte in vitro. Gamete Res 1985;12:29–39Google Scholar
  7. 7.
    Lambert H, Overstreet JW, Morales P, Hanson FW, Yanagimachi R: Sperm capacitation in the human female reproductive tract. Fertil Steril 1985;43:325–327Google Scholar
  8. 8.
    Cross NL, Lambert H, Samuels S: Sperm binding activity of the zona pellucida of immature mouse oocytes. Cell Biol Rep 1986;10:545–554Google Scholar
  9. 9.
    Yanagimachi R, Lopata A, Odom CB, Bronson RA, Mahi CA, Nicolson GL: Retention of biologic characteristics of zona pellucida in highly concentrated salt solution: The use of salt-stored eggs for assessing the fertilizing capacity of spermatozoa. Fertil Steril 1979;31:562–574Google Scholar
  10. 10.
    Burkman LJ, Coddington CC, Franken DR, Kruger TF, Rosenwaks Z, Hodgen GD: The hemizona assay (HZA): Development of a diagnostic test for the binding of human spermatozoa to human hemizona pellucida to predict fertilization potential. Fertil Steril 1988;49:688–697Google Scholar
  11. 11.
    Franken DR, Burkman LJ, Oehninger SC, Coddington CC, Veeck LL, Kruger TF, Rosenwaks Z, Hodgen GD: The hemizona assay using salt stored human oocytes: Evaluation of zona pellucida capacity for binding human spermatozoa. Gamete Res 22:15–26, 1989Google Scholar
  12. 12.
    Kruger TD, Menkveld R, Stander FSH, Lombard CJ, Van der Merwe JP, Van Zyl JA, Smith K: Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil Steril 1986;46:1118–1123Google Scholar
  13. 13.
    Kruger TF, Ackerman SB, Simmons KF, Swanson RJ, Brugo S, Acosta AA. A quick reliable staining techinque for sperm morphology. Arch Androl 1987;18:275–278Google Scholar
  14. 14.
    Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF, Oehninger S: Predictive value of abnormal sperm morphology in IVF. Fertil Steril 1988;49:112–117Google Scholar
  15. 15.
    Acosta AA, van Uem J, Ackerman SB, Mayer JF: Estimation of male fertility by examination and testing of spermatozoa.In In Vitro Fertilization-Norfolk. HW Jones Jr, GS Jones, HD Hodgen, Z Rosenwaks (eds). Baltimore, Williams and Wilkins, 1986, pp 126–161Google Scholar
  16. 16.
    Jones HW Jr, Jones GS, Hodgen GD, Rosenwaks Z (eds): In Vitro Fertilization-Norfolk. Baltimore, Williams and Wilkins, 1986Google Scholar
  17. 17.
    Veeck LL: Atlas of the Human Oocyte and Early Conceptus. Baltimore, Williams and Wilkins, 1986Google Scholar
  18. 18.
    Hofmann N, Haider SG: Neue ergebnisse morphologischer diagnostik der spermatogenesestorungen. Gynakologie 1985;18:70Google Scholar
  19. 19.
    Mathur S, Carlton M, Ziegler J, Rust PF, Williamson HO: A computerized sperm motion analysis. Fertil Steril 1986;46:484–488Google Scholar
  20. 20.
    Knuth UA, Yeung CH, Nieschlag E: Computerized semen analysis: Objective measurement of semen characteristics is biased by subjective parameter setting. Fertil Steril 1987;48:118–124Google Scholar
  21. 21.
    Remington RD, Schork MA: Statistics with Applications to the Biological and Health Sciences. Prentice-Hall, Englewood Cliffs, NJ, 1970, p 152Google Scholar
  22. 22.
    SPSSX User's Guide, 2nd ed. New York, McGraw-Hill, 1986Google Scholar
  23. 23.
    Chen C, Sathananthan AH: Early penetration of human sperm through the vestments of human eggs in vitro. Arch Androl 1986;16:183–197Google Scholar
  24. 24.
    Mahadevan MD, Trounson AO, Wood C, Leeton JF: Effect of oocyte quality and sperm characteristics on the number of spermatozoa bound to the zona pellucida of human oocytes inseminated in vitro. J Vitro Fert Embryo Transfer 1987;4:223–241Google Scholar
  25. 25.
    Rogers BJ, Bentwood BJ, Vancampen H, Helmbrecht G, Soderdal D, Hale RW: Sperm morphology assessment as an indicator of human fertilizing capacity. J Androl 1983;4:119Google Scholar
  26. 26.
    Testart J, Lassalle B, Frydman R, Belaisch JC: A study of factors affecting the success of human fertilization in vitro. II. Influence of semen quality and oocyte maturity on fertilization and cleavage. Biol Reprod 1983;28:425–431Google Scholar
  27. 27.
    Mahadevan MM, Trounson AO: The influence of seminal characteristics on the success rate of human in vitro fertilization. Fertil Steril 1984;42:400–405Google Scholar
  28. 28.
    Courtot AM, Escalier D, Jouannet P, David G: Impaired ability of human spermatozoa to penetrate zona-free hamster oocytes: Is a postacrosomal sheath anomaly involved? Gamete Res 1987;17:145–156Google Scholar

Copyright information

© Plenum Publishing Corporation 1989

Authors and Affiliations

  • Daniel R. Franken
    • 1
  • Sergio Oehninger
    • 4
  • Lani Johnson Burkman
    • 4
  • Charles C. Coddington
    • 2
  • Thinus F. Kruger
    • 3
  • Zev Rosenwaks
    • 4
  • Anibal A. Acosta
    • 4
  • Gary D. Hodgen
    • 4
  1. 1.Department of Obstetrics and Gynecology, Faculty of MedicineUniversity of the Organe Free StateBloemfonteinRepublic of South Africa
  2. 2.Department of Obstetrics and GynecologyPortsmouth Naval HospitalPortsmouth
  3. 3.Infertility ClinicTygerberg Hospital and University of StellenboschParow
  4. 4.The Jones I Institute for Reproductive Medicine, Department of Obstetrics and GynecologyEastern Virginia Medical SchoolNorfolk

Personalised recommendations