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Effect of apoptosis and reactive oxygen species productionin human granulosa cells on oocyte fertilizationand blastocyst development

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

Abstract

Purpose: The aim was to establish the impact of human granulosa cell apoptosis and reactive oxygen species (ROS) production on fertilization competence of the oocyte, embryo developmental stage and implantation rate.

Methods: Thirty women undergoing IVF-ET for tubal factor infertility were included; GnRH antagonists and gonadotrophins were used for ovarian stimulation. Granulosa cells were isolated from each aspirated follicle using gradient centrifugation. Apoptosis was studied by flow cytometry using annexin V and propidium iodide. ROS production was studied with hydroethidine staining and analyzed by flow cytometry.

Results: There were no differences in characteristics of granulosa cells between the follicles with fertilized and non-fertilized oocytes. The analyzed characteristics of granulosa cells in corresponding follicles had no effect on embryo developmental stage on day 5. The percentage of ROS producing granulosa cells was lower in the follicles giving rise to blastocysts that resulted in implantation compared to those that did not (39.9% versus 69.9%,P = 0.031).

Conclusions: Apoptosis and ROS production in granulosa cells have no significant impact on fertilization and do not correlate with the development of blastocysts. An increased percentage of ROS producing granulosa cells results in fewer oocytes retrieved and diminishes implantation rate.

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Acknowledgements

The authors wish to thank all members of our IVF team for assistance in the study, and Mojca Pirc, B.A., for editing the text.

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Authors and Affiliations

  1. Reproductive Unit, Department of Obstetrics and Gynecology, University Medical Centre Ljubljana, Šlajmerjeva 3, SI-1000, Ljubljana, Slovenia

    Nina Jančar, Irma Virant Klun & Eda Vrtačnik Bokal

  2. Laboratory for Lymphocytes and Cytometry, Institute of Microbiology and Immunology, Medical Faculty,University of Ljubljana, Korytkova 2, SI-1000, Ljubljana, Slovenia

    Andreja N. Kopitar & Alojz Ihan

Authors
  1. Nina Jančar

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  2. Andreja N. Kopitar

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  3. Alojz Ihan

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  4. Irma Virant Klun

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  5. Eda Vrtačnik Bokal

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Correspondence toNina Jančar.

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Jančar, N., Kopitar, A.N., Ihan, A.et al. Effect of apoptosis and reactive oxygen species productionin human granulosa cells on oocyte fertilizationand blastocyst development.J Assist Reprod Genet24, 91–97 (2007). https://doi.org/10.1007/s10815-006-9103-8

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