Pre-clinical validation of a closed surface system (Cryotop SC) for the vitrification of oocytes and embryos in the mouse model |
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| Institution: | 1. IVI Valencia, IVI-RMA Global, Plaza Policía Local, 3, 46015, Valencia, Spain;2. Embryotools, Parc Científic de Barcelona, Av. Doctor Marañon, 8, 08028, Barcelona, Spain;1. Embryo Technology and Stem Cell Research Center, Thailand;2. School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;3. School of Anatomy, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;1. Instituto Nacional de Investigação Agrária e Veterinária (National Institute of Agrarian and Veterinarian Research), Unidade de Biotecnologia e Recursos Genéticos, Vale de Santarém, Portugal;2. Research Institute for Medicines (iMed. ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Lisboa, Portugal;3. Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal;4. Centro de Investigação Interdisciplinar em Sanidade Animal (CIISA), Faculdade de Medicina Veterinária, Universidade de Lisboa, Lisboa, Portugal;5. Dpto. Bioquímica e Biologia Humana, Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal;1. Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan 650224, People''s Republic of China;2. College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, People''s Republic of China;1. College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, 471023, China;2. Henan Provincial Key Laboratory for Grass-Feeding Animal, Henan University of Science and Technology, Luoyang, 471023, China;1. Department of Mechanical Engineering, Villanova University, Villanova, PA 19085, USA;2. Institute of Molecular Medicine and Genetics, Department of Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA;3. School of Chemical, Biological and Environmental Engineering, Oregon State University, Corvallis, OR 97331, USA;4. Department of Obstetrics and Gynecology, and Cancer Center, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA |
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| Abstract: | Vitrification is currently a well-established technique for the cryopreservation of oocytes and embryos. It can be achieved either by direct (open systems) or indirect (closed systems) contact with liquid nitrogen. While there is not a direct evidence of disease transmission by transferred cryopreserved embryos, it was experimentally demonstrated that cross-contamination between liquid nitrogen and embryos may occur, and thus, the use of closed devices has been recommended to avoid the risk of contamination. Unfortunately, closed systems may result in lower cooling rates compared to open systems, due to the thermal insulation of the samples, which may cause ice crystal formation resulting in impaired results. In our study, we aimed to validate a newly developed vitrification device (Cryotop SC) that has been specifically designed for being used as a closed system. The cooling and warming rates calculated for the closed system were 5.254 °C/min and 43.522 °C/min, respectively. Results obtained with the closed system were equivalent to those with the classic Cryotop (open system), with survival rates in oocytes close to 100%. Similarly, the potential of the survived oocytes to develop up to good quality blastocysts after parthenogenetic activation between both groups was statistically equivalent. Assessment of the meiotic spindle and chromosome distribution by fluorescence microscopy in vitrified oocytes showed alike morphologies between the open and closed system. No differences were found either between the both systems in terms of survival rates of one-cell stage embryos or blastocysts, as well as, in the potential of the vitrified/warmed blastocysts to develop to full-term after transferred to surrogate females. |
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| Keywords: | Cryotop Vitrification Surface closed system |
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