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. 1999 Jan;16(1):24–29. doi: 10.1023/A:1022541511875

Use of Failed-Fertilized Oocytes for Diagnostic Zona Binding Purposes After Sperm Binding Improvement with a Modified Medium

Ralf Henkel 1,✉, Christina Müller 1, Thomas Stalf 2, Wolf-Bernhard Schill 1, Daniel R Franken 3
PMCID: PMC3468209  PMID: 9987690

Abstract

Purpose:Because the availability of prophase oocytes for zona binding testing is limited, we compared sperm binding to the zona of failed-fertilized intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) oocytes after incubation in a standard IVF medium and a specially composed binding improvement medium.

Methods:Semen samples from nine patients and nine fertile donors were separated in parallel by the standard swim-up method in both media. Subsequently, hemizona assays were performed with prophase, failed-fertilized ICSI and IVF oocytes.

Results:Sperm separation resulted in a significantly higher sperm count (P < 0.01) and progressive motility (P = 0.018) in binding improvement medium. Moreover, spermatozoa coincubated with hemizonae (prophase, failed-fertilized ICSI and IVF oocytes) in binding improvement medium bound significantly more to hemizonae than in the controls (P < 0.01). However, the hemizona index did not differ.

Conclusions:Thus, the limited number of human zonae can be increased by the use of oocytes that failed to fertilize during ICSI or IVF. This will lead to a qualitative improvement of the diagnostic spectrum in male-factor infertility.

Keywords: human sperm, zona pellucida, sperm binding, hemizona assay, failed-fertilized ICSI and IVF oocytes

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REFERENCES

  • 1.Oehninger S, Veeck L, Franken D, Kruger TF, Acosta AA, Hodgen GD. Human preovulatory oocytes have a higher sperm-binding ability than immature oocytes under hemizona assay conditions: Evidence supporting the concept of “zona maturation.”. Fertil Steril. 1991;55:1165–1170. doi: 10.1016/s0015-0282(16)54369-6. [DOI] [PubMed] [Google Scholar]
  • 2.Oehninger S, Acosta A, Veeck L, Brzyski R, Kruger TF, Muasher SJ, Hodgen GD. Recurrent failure of in vitro fertilization: Role of the hemizona assay in the sequential diagnosis of specific sperm-oocyte defects. Am J Obstet Gynecol. 1991;16:1210–1215. doi: 10.1016/0002-9378(91)90684-j. [DOI] [PubMed] [Google Scholar]
  • 3.Franken DR, Kruger TF, Oehninger S, Coddington CC, Lombard C, Smith K, Hodgen GD. The ability of the hemizona assay to predict human fertilization in different and consecutive in-vitro fertilization cycles. Hum Reprod. 1993;8:1240–1244. doi: 10.1093/oxfordjournals.humrep.a138234. [DOI] [PubMed] [Google Scholar]
  • 4.Coddington CC, Oehninger S, Olive DL, Franken DR, Kruger TF, Hodgen GD. Hemizona index (HZI) demonstrates excellent predictability when evaluating sperm fertilizing capacity in in vitro fertilization patients. J Androl. 1994;15:250–254. [PubMed] [Google Scholar]
  • 5.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 the human hemizona pellucida to predict fertilization potential. Fertil Steril. 1988;49:688–697. [PubMed] [Google Scholar]
  • 6.Familiari G, Nottola SA, Micara G, Aragona C, Motta PM. Is the sperm-binding capability of the zona pellucida linked to its surface structure? A scanning electron microscopic study of human in vitro fertilization. J In Vitro Fert Embryo Transfer. 1988;5:134–143. doi: 10.1007/BF01131175. [DOI] [PubMed] [Google Scholar]
  • 7.Tesarik J, Pilka L, Travnik P. Zona pellucida resistance to sperm penetration before the completion of human oocyte maturation. J Reprod Fertil. 1988;83:483–495. doi: 10.1530/jrf.0.0830487. [DOI] [PubMed] [Google Scholar]
  • 8.Henkel R, Cooper S, Kaskar K, Schill W-B, Habenicht U-F, Franken DR. Influence of elevated pH levels on structural and functional characteristics of the human zona pellucida: Functional morphological aspects. J Assist Reprod Genet. 1995;12:644–649. doi: 10.1007/BF02212590. [DOI] [PubMed] [Google Scholar]
  • 9.Wolf DP. The mammalian egg's block to polyspermy. In: Mastroianni L, Biggers JD, editors. Fertilization and Embryonic Development in Vitro. New York: Raven Press; 1981. pp. 183–197. [Google Scholar]
  • 10.Franken DR, Windt ML, Kruger TF, Oehninger S, Hodgen GD. Comparison of sperm binding potential of uninseminated, inseminated-unfertilized, and fertilized-noncleaved human oocytes under hemizona assay conditions. Mol Reprod Dev. 1991;30:56–61. doi: 10.1002/mrd.1080300108. [DOI] [PubMed] [Google Scholar]
  • 11.Franken DR, Coddington CC, Burkman LJ, Oosthuizen WT, Oehninger SC, Kruger TF, Hodgen GD. Defining the valid hemizona assay: Accounting for binding variability within zonae pellucidae and within semen samples from fertile males. Fertil Steril. 1991;56:1156–1161. [PubMed] [Google Scholar]
  • 12.Quinn P, Kerin JF, Warnes GM. Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid. Fertil Steril. 1985;44:493–498. doi: 10.1016/s0015-0282(16)48918-1. [DOI] [PubMed] [Google Scholar]
  • 13.Shabanowitz RB, O'Rand MG. Molecular changes in the human zona pellucida associated with fertilization and human sperm-zona interactions. Ann NY Acad Sci. 1988;541:621–632. doi: 10.1111/j.1749-6632.1988.tb22299.x. [DOI] [PubMed] [Google Scholar]
  • 14.Okamura N, Tajima Y, Soejima A, Masuda H, Sugita Y. Sodium bicarbonate in seminal plasma stimulates the motility of mammalian spermatozoa through direct activation of adenylate cyclase. J Biol Chem. 1985;260:9699–9705. [PubMed] [Google Scholar]
  • 15.Tajima Y, Okamura N, Sugita Y. The activating effects of bicarbonate on sperm motility and respiration at ejaculation. Biochim Biophys Acta. 1987;924:519–529. doi: 10.1016/0304-4165(87)90168-1. [DOI] [PubMed] [Google Scholar]
  • 16.Boatman DE, Robbins RS. Bicarbonate: carbon-dioxide regulation of sperm capacitation, hyperactivated motility, and acrosome reactions. Biol Reprod. 1991;44:806–813. doi: 10.1095/biolreprod44.5.806. [DOI] [PubMed] [Google Scholar]
  • 17.Si Y, Okuno M. Multiple activation of mouse sperm motility. Mol Reprod Dev. 1993;36:89–95. doi: 10.1002/mrd.1080360113. [DOI] [PubMed] [Google Scholar]
  • 18.Parkkila S, Rajaniemi H, Kellokumpu S. Polarized expression of a band 3-related protein in mammalian sperm cells. Biol Reprod. 1993;49:326–331. doi: 10.1095/biolreprod49.2.326. [DOI] [PubMed] [Google Scholar]
  • 19.Si Y, Okuno M. Activation of mammalian sperm motility by regulation of microtubule sliding via cyclic adenosine 5'-monophosphate-dependent phosphorylation. Biol Reprod. 1995;53:1081–1087. doi: 10.1095/biolreprod53.5.1081. [DOI] [PubMed] [Google Scholar]
  • 20.Stauss CR, Votta TJ, Suarez SS. Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida. Biol Reprod. 1995;53:1280–1285. doi: 10.1095/biolreprod53.6.1280. [DOI] [PubMed] [Google Scholar]
  • 21.Henkel R, Franken DR, Habenicht U-F. Zona pellucida as physiological trigger for the induction of acrosome reaction. Andrologia. 1998;30:275–280. doi: 10.1111/j.1439-0272.1998.tb01171.x. [DOI] [PubMed] [Google Scholar]
  • 22.Coddington CC, Fulgham DL, Alexander NJ, Johnson D, Herr JC, Hodgen GD. Sperm bound to zona pellucida in hemizona assay demonstrate acrosome reaction when stained with T-6 antibody. Fertil Steril. 1990;54:504–508. [PubMed] [Google Scholar]
  • 23.Franken DR, Oosthuizen WT, Cooper S, Kruger TF, Burkman LJ, Coddington CC, Hodgen GD. Electron microscopic evidence on the acrosomal status of bound sperm and their penetration into human hemizonae pellucida after storage in a buffered salt solution. Andrologia. 1991;23:205–208. doi: 10.1111/j.1439-0272.1991.tb02538.x. [DOI] [PubMed] [Google Scholar]
  • 24.Veeck LL, Wortham JWE, Jr, Witmyer J, Sandow BA, Acosta AA, Garcia JE, Jones GS, Jones HW., Jr Maturation and fertilization of morphologically immature human oocytes in a program of in vitro fertilization. Fertil Steril. 1983;39:594–602. doi: 10.1016/s0015-0282(16)47052-4. [DOI] [PubMed] [Google Scholar]
  • 25.Winston NJ, Braude PR, Johnson MH. Are failed-fertilized human oocytes useful? Hum Reprod. 1993;8:503–507. doi: 10.1093/oxfordjournals.humrep.a138084. [DOI] [PubMed] [Google Scholar]
  • 26.Nagy ZP, Joris H, Liu J, Staessen C, Devroey P, Van Steirteghem AC. Intracytoplasmic single sperm injection of 1-day-old unfertilized human oocytes. Hum Reprod. 1993;8:2180–2184. doi: 10.1093/oxfordjournals.humrep.a138000. [DOI] [PubMed] [Google Scholar]
  • 27.Khan I, Staessen C, Van den Abbeel E, Camus M, Wisanto A, Smitz J, Devroey P, Van Steirteghem AC. Time of insemination and its effect on in vitro fertilization, cleavage and pregnancy rates in GnRH agonist/HMG-stimulated cycles. Hum Reprod. 1989;4:921–926. doi: 10.1093/oxfordjournals.humrep.a137013. [DOI] [PubMed] [Google Scholar]

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