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The combination of calcium ionophore A23187 and GM-CSF can safely salvage aged human unfertilized oocytes after ICSI

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Artificial oocyte activation using calcium ionophores and enhancement of embryonic developmental potential by the granulocyte-macrophage colony-stimulating factor (GM-CSF) have already been reported. In this study, we evaluated the synergistic effect of these two methods on aged human unfertilized oocytes after intracytoplasmic sperm injection (ICSI). Then, we cultured the resulting embryos to the blastocyst stage and screened them for chromosomal abnormalities, to assess the safety of this protocol.

Methods

Aged human oocytes deemed unfertilized after ICSI were activated, either by briefly applying the calcium ionophore A23187 alone (group A) or by briefly applying the ionophore and then supplementing the culture medium with recombinant human GM-CSF (rhGM-CSF) (group B). Next, the development was monitored in a time-lapse incubator system, and ploidy was analyzed by array comparative genomic hybridization (aCGH), after whole embryo biopsy and whole genome amplification. Differences between oocytes and resulting embryos in both groups were evaluated statistically.

Results

Oocytes unfertilized after ICSI can be activated with the calcium ionophore A23187 to show two pronuclei and two polar bodies. Addition of rhGM-CSF in the culture medium of A23187-activated oocytes enhances their cleaving and blastulation potential and results in more euploid blastocysts compared to the culture medium alone.

Conclusions

This study shows that activating post-ICSI aged human unfertilized oocytes with a combination of a calcium ionophore and a cytokine can produce good-morphology euploid blastocysts.

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Acknowledgments

The authors wish to thank Mr. Christos Karamalegos for the help with statistical analysis and Ms. Maria Papadopoulou for collecting the patient consent forms.

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Authors and Affiliations

  1. Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece

    Konstantinos A. Economou, Dimitra Christopikou, Erika Tsorva, Stephen Davies, Minas Mastrominas & Haris Cazlaris

  2. Department of Physiology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece

    Michael Koutsilieris & Panagoula Angelogianni

  3. First Department of Obstetrics and Gynaecology, School of Medicine, Alexandra University Hospital, National and Kapodistrian University of Athens, Athens, Greece

    Dimitris Loutradis

Authors
  1. Konstantinos A. Economou
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  2. Dimitra Christopikou
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  3. Erika Tsorva
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  4. Stephen Davies
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  5. Minas Mastrominas
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  6. Haris Cazlaris
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  7. Michael Koutsilieris
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  9. Dimitris Loutradis
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Corresponding author

Correspondence to Konstantinos A. Economou.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

The study was approved by the institutional research ethics committee, and then it was submitted to and obtained authorization by the Hellenic National Authority of Medically Assisted Reproduction (authorization ref. 134/15), as required by national legislation for research use of human embryos not destined to embryo transfer.

Informed consent

All patients who donated oocytes and embryos to be included in the study signed consent forms, after receiving exhaustive information on the purpose of the research carried out, on the procedures to be used, on the possibility to withdraw their consent at any time up to the first actual use of each oocyte or embryo in an experiment, and on the legal and ethical obligation of the authors to destroy all embryos after completion of the experiments, without attempting to transfer them in vivo.

Additional information

Capsule The exposure of human unfertilized oocytes 18 h after ICSI to a combination of calcium ionophore A23187 and the GM-CSF cytokine can safely salvage the oocytes, resulting in the development of good-morphology, euploid blastocysts, with potential clinical use.

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Economou, K.A., Christopikou, D., Tsorva, E. et al. The combination of calcium ionophore A23187 and GM-CSF can safely salvage aged human unfertilized oocytes after ICSI. J Assist Reprod Genet 34, 33–41 (2017). https://doi.org/10.1007/s10815-016-0823-0

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