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Validation of the hemizona assay (HZA) in a monkey model. II. Kinetics of binding and influence of cryopreserved—thawed spermatozoa

  • Sergio Oehninger
  • Mahmood Morshedi
  • Hakan Ertunc
  • Christine Philput
  • Silvina M. Bocca
  • Anibal A. Acosta
  • Gary D. Hodgen
Animal Investigations

Abstract

We compared fresh and frozen—thawed cynomolgus monkey spermatozoa tight binding to the zona pellucida under hemizona assay (HZA) conditions. Monkey oocytes were recovered after superovulation and stored in salt solution. Matching hemizonae were obtained by micromanipulation. Semen, obtained by electroejaculation, was used fresh or was cyropreserved, thawed, and washed by swim-up separation. At the standard initial dilution of 500,000 motile sperm/ml (or 5 × 104 motile sperm/hemizona), binding was significantly higher for fresh sperm (P = 0.00004). For frozen—thawed samples, there was a linear increase in the number of tightly bound sperm with increasing sperm concentration (r = 0.95). At 1.5 × 106 motile sperm/hemizona, binding of frozen—thawed spermatozoa was similar to that of fresh at a standard concentration. Kinetic studies showed peak binding at 1 hr of gametes coincubation. We conclude that, in this monkey model, the HZA is a valuable bioassay for evaluation of sperm binding to the zona pellucida, the initial requisite for fertilization and embryo development.

Key words

sperm zona pellucida binding hemizona assay 

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Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Sergio Oehninger
    • 1
  • Mahmood Morshedi
    • 1
  • Hakan Ertunc
    • 1
  • Christine Philput
    • 2
  • Silvina M. Bocca
    • 1
  • Anibal A. Acosta
    • 1
  • Gary D. Hodgen
    • 1
  1. 1.The Jones Institute for Reproductive Medicine, Department of Obstetrics and GynecologyEastern Virginia Medical SchoolNorfolkUSA
  2. 2.Department of Radiation Oncology (Biostatistics)Eastern Virginia Medical SchoolNorfolkUSA

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