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Pre-Implantation Mouse Embryos Cultured In Vitro under Different Oxygen Concentrations Show Altered Ultrastructures

Author

Listed:
  • Manuel Belli

    (Department of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, Italy)

  • Paolo Rinaudo

    (Center for Reproductive Sciences, Department of Obgyn, University of California San Francisco, San Francisco, CA 94143, USA)

  • Maria Grazia Palmerini

    (Department of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, Italy)

  • Elena Ruggeri

    (Center for Reproductive Sciences, Department of Obgyn, University of California San Francisco, San Francisco, CA 94143, USA)

  • Sevastiani Antonouli

    (Department of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, Italy)

  • Stefania Annarita Nottola

    (Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, La Sapienza University of Rome, 00161 Rome, Italy)

  • Guido Macchiarelli

    (Department of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, Italy)

Abstract

Assisted Reproductive Technologies routinely utilize different culture media and oxygen (O 2 ) concentrations to culture human embryos. Overall, embryos cultured under physiological O 2 tension (5%) have improved development compared to embryos cultured under atmospheric O 2 conditions (20%). The mechanisms responsible for this remain unclear. This study aimed to evaluate the effect of physiologic (5%) or atmospheric O 2 (20%) tension on the microscopic ultrastructure of pre-implantation mouse embryos using Transmission Electron Microscopy (TEM). Embryos flushed out of the uterus after natural mating were used as the control. For use as the control, 2-cells, 4-cells, morulae, and blastocysts were flushed out of the uterus after natural fertilization. In vitro fertilization (IVF) was performed using potassium simplex optimized medium (KSOM) under different O 2 tensions (5% and 20%) until the blastocyst stage. After collection, embryos were subjected to the standard preparative for light microscopy (LM) and TEM. We found that culture in vitro under 5% and 20% O 2 results in an increase of vacuolated shaped mitochondria, cytoplasmic vacuolization and presence of multi-vesicular bodies at every embryonic stage. In addition, blastocysts generated by IVF under 5% and 20% O 2 showed a lower content of heterochromatin, an interruption of the trophectodermal and inner cell mass cell membranes, an increased density of residual bodies, and high levels of glycogen granules in the cytoplasm. In conclusion, this study suggests that in vitro culture, particularly under atmospheric O 2 tension, causes stage-specific changes in preimplantation embryo ultrastructure. In addition, atmospheric (20%) O 2 is associated with increased alterations in embryonic ultrastructure; these changes may explain the reduced embryonic development of embryos cultured with 20% O 2 .

Suggested Citation

  • Manuel Belli & Paolo Rinaudo & Maria Grazia Palmerini & Elena Ruggeri & Sevastiani Antonouli & Stefania Annarita Nottola & Guido Macchiarelli, 2020. "Pre-Implantation Mouse Embryos Cultured In Vitro under Different Oxygen Concentrations Show Altered Ultrastructures," IJERPH, MDPI, vol. 17(10), pages 1-17, May.
    • Handle: RePEc:gam:jijerp:v:17:y:2020:i:10:p:3384-:d:357315
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    References listed on IDEAS

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    1. Serena Bianchi & Stefania Annarita Nottola & Diana Torge & Maria Grazia Palmerini & Stefano Necozione & Guido Macchiarelli, 2020. "Association between Female Reproductive Health and Mancozeb: Systematic Review of Experimental Models," IJERPH, MDPI, vol. 17(7), pages 1-15, April.
    2. Wolf Reik, 2007. "Stability and flexibility of epigenetic gene regulation in mammalian development," Nature, Nature, vol. 447(7143), pages 425-432, May.
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