Stepped vitrification technique for human ovarian tissue cryopreservation
Autor(a) principal: | |
---|---|
Data de Publicação: | 2019 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1038/s41598-019-56585-7 http://hdl.handle.net/11449/201434 |
Resumo: | 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. |
id |
UNSP_822545c452948e78486761204b2f99a1 |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/201434 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Stepped vitrification technique for human ovarian tissue cryopreservationThe 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.Fonds De La Recherche Scientifique - FNRSPôle de Recherche en Gynécologie Institut de Recherche Expérimentale et Clinique Université Catholique de Louvain, Avenue Mounier 52, bte B1.52.02Departament of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Rua Cristóvão Colombo 2265 Jardim NazarethCentro Nacional de Aceleradores (CNA) University of Seville, Calle Thomas Alva Edison 7Engineering School of Sevilla University of Seville Camino Descubrimientos S/N Isla CartujaService d’Anatomie Pathologique Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10General Electric Healthcare Sovereign HouseGynecology and Andrology Department Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10Departament of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Rua Cristóvão Colombo 2265 Jardim NazarethFonds De La Recherche Scientifique - FNRS: 5/4/150/5Université Catholique de LouvainUniversidade Estadual Paulista (Unesp)University of SevilleIsla CartujaCliniques Universitaires Saint-LucSovereign HouseLeonel, Ellen Cristina Rivas [UNESP]Corral, AriadnaRisco, RamonCamboni, AlessandraTaboga, Sebastião Roberto [UNESP]Kilbride, PeterVazquez, MarinaMorris, JohnDolmans, Marie-MadeleineAmorim, Christiani A.2020-12-12T02:32:27Z2020-12-12T02:32:27Z2019-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1038/s41598-019-56585-7Scientific Reports, v. 9, n. 1, 2019.2045-2322http://hdl.handle.net/11449/20143410.1038/s41598-019-56585-72-s2.0-85077169147Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengScientific Reportsinfo:eu-repo/semantics/openAccess2021-10-22T19:03:43Zoai:repositorio.unesp.br:11449/201434Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T23:21:02.788112Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Stepped vitrification technique for human ovarian tissue cryopreservation |
title |
Stepped vitrification technique for human ovarian tissue cryopreservation |
spellingShingle |
Stepped vitrification technique for human ovarian tissue cryopreservation Leonel, Ellen Cristina Rivas [UNESP] |
title_short |
Stepped vitrification technique for human ovarian tissue cryopreservation |
title_full |
Stepped vitrification technique for human ovarian tissue cryopreservation |
title_fullStr |
Stepped vitrification technique for human ovarian tissue cryopreservation |
title_full_unstemmed |
Stepped vitrification technique for human ovarian tissue cryopreservation |
title_sort |
Stepped vitrification technique for human ovarian tissue cryopreservation |
author |
Leonel, Ellen Cristina Rivas [UNESP] |
author_facet |
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. |
author_role |
author |
author2 |
Corral, Ariadna Risco, Ramon Camboni, Alessandra Taboga, Sebastião Roberto [UNESP] Kilbride, Peter Vazquez, Marina Morris, John Dolmans, Marie-Madeleine Amorim, Christiani A. |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Université Catholique de Louvain Universidade Estadual Paulista (Unesp) University of Seville Isla Cartuja Cliniques Universitaires Saint-Luc Sovereign House |
dc.contributor.author.fl_str_mv |
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. |
description |
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. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-12-01 2020-12-12T02:32:27Z 2020-12-12T02:32:27Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1038/s41598-019-56585-7 Scientific Reports, v. 9, n. 1, 2019. 2045-2322 http://hdl.handle.net/11449/201434 10.1038/s41598-019-56585-7 2-s2.0-85077169147 |
url |
http://dx.doi.org/10.1038/s41598-019-56585-7 http://hdl.handle.net/11449/201434 |
identifier_str_mv |
Scientific Reports, v. 9, n. 1, 2019. 2045-2322 10.1038/s41598-019-56585-7 2-s2.0-85077169147 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Scientific Reports |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129509085413376 |