Stefania Annarita Nottola1, Elena Albani2, Giovanni Coticchio3,4, Maria Grazia Palmerini5, Caterina Lorenzo5, Giulia Scaravelli6, Andrea Borini3, Paolo Emanuele Levi-Setti2, Guido Macchiarelli5. 1. Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, Sapienza University, Rome, Italy. stefania.nottola@uniroma1.it. 2. Humanitas Research Hospital, Department of Gynecology, Division of Gynecology and Reproductive Medicine, Humanitas Fertility Center, Rozzano, Milan, Italy. 3. Tecnobios Procreazione, Centre for Reproductive Health, Bologna, Italy. 4. Biogenesi, Reproductive Medicine Centre, Monza, Italy. 5. Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy. 6. CNESPS, National Health Institute, Rome, Italy.
Abstract
PURPOSE: Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. METHODS: Samples were studied by light and transmission electron microscopy. RESULTS: We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. CONCLUSIONS: This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism.
PURPOSE: Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. METHODS: Samples were studied by light and transmission electron microscopy. RESULTS: We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. CONCLUSIONS: This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism.
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