Autor(es): Leonel, Ellen Cristina Rivas [UNESP] ; Corral, Ariadna ; Risco, Ramon ; Camboni, Alessandra ; Taboga, Sebastião Roberto [UNESP] ; Kilbride, Peter ; Vazquez, Marina ; Morris, John ; Dolmans, Marie-Madeleine ; Amorim, Christiani A.
Data: 2020
Identificador Persistente: http://hdl.handle.net/11449/201434
Origem: Oasisbr
Made available in DSpace on 2020-12-12T02:32:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01
Fonds De La Recherche Scientifique - FNRS
The advantage of stepped vitrification (SV) is avoiding ice crystal nucleation, while decreasing the toxic effects of high cryoprotectant concentrations. We aimed to test this method for human ovarian tissue cryopreservation. Ovarian cortex was taken from 7 fertile adult women. Samples were subjected to an SV protocol performed in an automatic freezer, which allowed sample transfer to ever higher concentrations of dimethyl sulfoxide (DMSO) as the temperature was reduced. Histological evaluation of the vitrified-warmed tissue showed large numbers of degenerated follicles after 24 hours of in vitro culture. We therefore evaluated DMSO perfusion rates by X-ray computed tomography, ice crystal formation by freeze-substitution, and cell toxicity by transmission electron microscopy, seeking possible reasons why follicles degenerated. Although cryoprotectant perfusion was considered normal and no ice crystals were formed in the tissue, ultrastructural analysis detected typical signs of DMSO toxicity, such as mitochondria degeneration, alterations in chromatin condensation, cell vacuolization and extracellular matrix swelling in both stromal and follicular cells. The findings indicated that the method failed to preserve follicles due to the high concentrations of DMSO used. However, adaptations can be made to avoid toxicity to follicles caused by elevated levels of cryoprotectants.
Pôle de Recherche en Gynécologie Institut de Recherche Expérimentale et Clinique Université Catholique de Louvain, Avenue Mounier 52, bte B1.52.02
Departament of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Rua Cristóvão Colombo 2265 Jardim Nazareth
Centro Nacional de Aceleradores (CNA) University of Seville, Calle Thomas Alva Edison 7
Engineering School of Sevilla University of Seville Camino Descubrimientos S/N Isla Cartuja
Service d’Anatomie Pathologique Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10
General Electric Healthcare Sovereign House
Gynecology and Andrology Department Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10
Departament of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Rua Cristóvão Colombo 2265 Jardim Nazareth
Fonds De La Recherche Scientifique - FNRS: 5/4/150/5
