Ultrastructure of immature and mature human oocytes after cryotop vitrification

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  • PALMERINI Maria Grazia
    Department of Life, Health and Environmental Sciences, University of L’Aquila, L’Aquila, Italy
  • ANTINORI Monica
    RAPRUI Day Hospital (International Associated Research Institute for Human Reproduction), Rome, Italy
  • MAIONE Marta
    Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, University La Sapienza, Rome, Italy
  • CERUSICO Fabrizio
    RAPRUI Day Hospital (International Associated Research Institute for Human Reproduction), Rome, Italy
  • VERSACI Caterina
    RAPRUI Day Hospital (International Associated Research Institute for Human Reproduction), Rome, Italy
  • NOTTOLA Stefania Annarita
    Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, University La Sapienza, Rome, Italy
  • MACCHIARELLI Guido
    Department of Life, Health and Environmental Sciences, University of L’Aquila, L’Aquila, Italy
  • KHALILI Mohammad Ali
    Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
  • ANTINORI Severino
    RAPRUI Day Hospital (International Associated Research Institute for Human Reproduction), Rome, Italy

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In vitro maturation of vitrified immature germinal vesicle (GV) oocytes is a promising fertility preservation option. We analyzed the ultrastructure of human GV oocytes after Cryotop vitrification (GVv) and compared it with fresh GV (GVc), fresh mature metaphase II (MIIc) and Cryotop-vitrified mature (MIIv) oocytes. By phase contrast microscopy and light microscopy, the oolemmal and cytoplasmic organization of fresh and vitrified oocytes did not show significant changes. GVv oocytes showed significant ultrastructural alterations of the microvilli in 40% of the samples; small vacuoles and occasional large/isolated vacuoles were abnormally present in the ooplasm periphery of 50% of samples. The ultrastructure of nuclei and mitochondria-vesicle (MV) complexes, as well as the distribution and characteristics of cortical granules (CGs), were comparable with those of GVc oocytes. MIIv oocytes showed an abnormal ultrastructure of microvilli in 30% of the samples and isolated large vacuoles in 70% of the samples. MV complexes were normal, but mitochondria-smooth endoplasmic reticulum aggregates appeared to be of reduced size. CGs were normally located under the oolemma but presented abnormalities in distribution and matrix electron density. In conclusion, Cryotop vitrification preserved main oocyte characteristics in the GV and MII stages, even if peculiar ultrastructural alterations appeared in both stages. This study also showed that the GV stage appears more suitable for vitrification than the MII stage, as indicated by the good ultrastructural preservation of important structures that are present only in immature oocytes, like the nucleus and migrating CGs.

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