Ovarian tissue cryopreservation by stepped vitrification and monitored by X-ray computed tomography |
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| Institution: | 1. Centro Nacional de Aceleradores (Universidad de Sevilla-CSIC-Junta de Andalucía), Calle Thomas Alva Edison 7, 41092, Sevilla, Spain;2. Departamento de Física Aplicada III, Escuela Técnica Superior de Ingeniería, Universidad de Sevilla, Camino Descubrimientos s/n, Isla Cartuja, 41092, Sevilla, Spain;3. Ginemed Clínicas Sevilla, Calle Farmaceutico Murillo Herrera 3, 41010, Sevilla, Spain;4. Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Avenue Mounier 52, Bte. B1.52.02, 1200, Brussels, Belgium;5. Cliniques Universitaires Saint-Luc, Gynecology Department, Avenue Hippocrate 10, 1200, Brussels, Belgium;6. Asymptote Limited, GE Healthcare, Sovereign House, Chivers Way, Vision Park, Cambridge, CB24 9ZR, United Kingdom;1. Center for Reproductive Medicine, Hospital for Maternity and Child Care of Linyi City, China;2. Nursing department, People''s Hospital of Linyi City, China;3. Department of Obstetrics & Gynecology, People''s Hospital of Laiwu City, China;4. Department of Reproductive Medicine, People''s Hospital of Laiwu City, China;1. School of Electrical Engineering and Computer Science, Louisiana State University, Baton Rouge, LA, 70803, USA;2. Center for Advanced Microstructures and Devices, Louisiana State University, Baton Rouge, LA, 70803, USA;3. Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA, 70820, USA;1. SRI, Société de Recherche pour l''Infertilité, Avenue Grandchamp 143, B-1150 Brussels, Belgium;2. Pôle de Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, & Cliniques Universitaires Saint-Luc, B-1200 Brussels, Belgium;1. Division of Orthopaedic Surgery, Department of Surgery, University of Alberta, Edmonton, Canada;2. Department of Chemical and Material Engineering, University of Alberta, Edmonton, Canada;3. Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Canada;4. Rabigh Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia |
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| Abstract: | Ovarian tissue cryopreservation is, in most cases, the only fertility preservation option available for female patients soon to undergo gonadotoxic treatment. To date, cryopreservation of ovarian tissue has been carried out by both traditional slow freezing method and vitrification, but even with the best techniques, there is still a considerable loss of follicle viability. In this report, we investigated a stepped cryopreservation procedure which combines features of slow cooling and vitrification (hereafter called stepped vitrification). Bovine ovarian tissue was used as a tissue model. Stepwise increments of the Me2SO concentration coupled with stepwise drops-in temperature in a device specifically designed for this purpose and X-ray computed tomography were combined to investigate loading times at each step, by monitoring the attenuation of the radiation proportional to Me2SO permeation. Viability analysis was performed in warmed tissues by immunohistochemistry. Although further viability tests should be conducted after transplantation, preliminary results are very promising. Four protocols were explored. Two of them showed a poor permeation of the vitrification solution (P1 and P2). The other two (P3 and P4), with higher permeation, were studied in deeper detail. Out of these two protocols, P4, with a longer permeation time at ?40 °C, showed the same histological integrity after warming as fresh controls. |
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| Keywords: | Slow vitrification X-ray computed tomography Ovarian tissue Dimethyl sulfoxide Preantral follicle |
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