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Fertilization failure and oocyte activation.

S Yamano1, K Nakagawa, H Nakasaka

  • 1Center for Maternity and Perinatal Care, University of Tokushima School of Medicine, Japan.

The Journal of Medical Investigation : JMI
|March 31, 2000
PubMed
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A novel method using calcium ionophore A23187 and puromycin effectively activates human unfertilized oocytes, producing parthenotes with normal karyotypes after intracytoplasmic sperm injection (ICSI). Further research is needed to assess safety for clinical use.

Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Assisted Reproductive Technology

Background:

  • Human oocytes are resistant to common activation methods like ethanol or calcium ionophore.
  • Existing methods like puromycin yield high activation rates but often result in abnormal karyotypes.
  • A method is needed to produce parthenotes with a single pronucleus and extruded second polar body for normal karyotype development.

Purpose of the Study:

  • To develop and evaluate a new oocyte activation method for human unfertilized oocytes.
  • To achieve parthenogenesis with a single pronucleus and second polar body extrusion.
  • To assess the potential for generating embryos with normal karyotypes.

Main Methods:

  • Utilized a combination of calcium ionophore A23187 and puromycin for oocyte activation.

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  • Applied the method to aged human and young mouse oocytes.
  • Tested the method on unfertilized human oocytes post-intracytoplasmic sperm injection (ICSI).

    • Main Results:

    • Achieved approximately 90% activation rate in aged human and young mouse oocytes.
    • Obtained about 80% parthenotes with a single pronucleus and extruded second polar body in aged human and young mouse oocytes.
    • Activated 30% of human unfertilized oocytes post-ICSI, resulting in two pronuclei with second polar body extrusion; four cleaved embryos showed normal karyotypes.

    Conclusions:

    • The combined calcium ionophore A23187 and puromycin method shows promise for activating human oocytes and producing parthenotes with normal karyotypes.
    • The method achieved high activation rates and a significant proportion of desired parthenotes.
    • Clinical application requires further investigation into the cytotoxic, teratogenetic, and mutagenetic effects of the agents used.