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Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice

OBJECTIVE: The aim of this study was to investigate the efficacy of protocols for mice ovary cryopreservation to compare the differences in Mouse Vasa Homologue expression (a germline cell marker) and ovarian viability after vitrification or slow freezing. METHODS: Female CF1 mice aged 40-45 days we...

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Autores principales: Terraciano, Paula Barros, Garcez, Tuane Alves, Berger, Markus, Durli, Isabel, Kuhl, Cristiana Palma, Batista, Vitória de Oliveira, Schneider, Raquel de Almeida, Festa, Jaquelline, Pilar, Emily, Goulart, Marcela, Ferreira, Charles, Passos, Eduardo Pandolfi, Lima, Elizabeth Cirne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Brazilian Society of Assisted Reproduction 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6993165/
https://www.ncbi.nlm.nih.gov/pubmed/31689043
http://dx.doi.org/10.5935/1518-0557.20190057
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author Terraciano, Paula Barros
Garcez, Tuane Alves
Berger, Markus
Durli, Isabel
Kuhl, Cristiana Palma
Batista, Vitória de Oliveira
Schneider, Raquel de Almeida
Festa, Jaquelline
Pilar, Emily
Goulart, Marcela
Ferreira, Charles
Passos, Eduardo Pandolfi
Lima, Elizabeth Cirne
author_facet Terraciano, Paula Barros
Garcez, Tuane Alves
Berger, Markus
Durli, Isabel
Kuhl, Cristiana Palma
Batista, Vitória de Oliveira
Schneider, Raquel de Almeida
Festa, Jaquelline
Pilar, Emily
Goulart, Marcela
Ferreira, Charles
Passos, Eduardo Pandolfi
Lima, Elizabeth Cirne
author_sort Terraciano, Paula Barros
collection PubMed
description OBJECTIVE: The aim of this study was to investigate the efficacy of protocols for mice ovary cryopreservation to compare the differences in Mouse Vasa Homologue expression (a germline cell marker) and ovarian viability after vitrification or slow freezing. METHODS: Female CF1 mice aged 40-45 days were randomly divided into three groups: Control, vitrification or slow freezing. Their ovaries were surgically removed, rinsed in saline solution and cryopreserved. For vitrification, we used a commercial protocol and for slow freeze, we used 1.5 M ethylene glycol (EG) as cryoprotectant. After that, the ovaries were processed for histological an immunohistochemical analysis, and counting of primordial, primary, pre-antral and antral follicles. RESULTS: No significant difference was found in the proportion of high-quality primordial, primary and pre-antral follicles after thawing/warming in the slow freezing and vitrification groups. The immunohistochemistry for MVH antibody demonstrated that the slow freeze group had a higher number of unmarked cells (p=0.012), indicating a harmful effect on the MVH expression in the ovarian tissue, where the cell structure is complex. CONCLUSION: Although both protocols indicated similar results in the histological analysis of follicular counts, the vitrification protocol was significantly better to preserve ovarian stem cells, an immature germ cell population. These cells are able to self-renew having regeneration potential, and may be effective for the treatment of ovarian failure and consequently infertility.
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spelling pubmed-69931652020-02-11 Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice Terraciano, Paula Barros Garcez, Tuane Alves Berger, Markus Durli, Isabel Kuhl, Cristiana Palma Batista, Vitória de Oliveira Schneider, Raquel de Almeida Festa, Jaquelline Pilar, Emily Goulart, Marcela Ferreira, Charles Passos, Eduardo Pandolfi Lima, Elizabeth Cirne JBRA Assist Reprod Original Article OBJECTIVE: The aim of this study was to investigate the efficacy of protocols for mice ovary cryopreservation to compare the differences in Mouse Vasa Homologue expression (a germline cell marker) and ovarian viability after vitrification or slow freezing. METHODS: Female CF1 mice aged 40-45 days were randomly divided into three groups: Control, vitrification or slow freezing. Their ovaries were surgically removed, rinsed in saline solution and cryopreserved. For vitrification, we used a commercial protocol and for slow freeze, we used 1.5 M ethylene glycol (EG) as cryoprotectant. After that, the ovaries were processed for histological an immunohistochemical analysis, and counting of primordial, primary, pre-antral and antral follicles. RESULTS: No significant difference was found in the proportion of high-quality primordial, primary and pre-antral follicles after thawing/warming in the slow freezing and vitrification groups. The immunohistochemistry for MVH antibody demonstrated that the slow freeze group had a higher number of unmarked cells (p=0.012), indicating a harmful effect on the MVH expression in the ovarian tissue, where the cell structure is complex. CONCLUSION: Although both protocols indicated similar results in the histological analysis of follicular counts, the vitrification protocol was significantly better to preserve ovarian stem cells, an immature germ cell population. These cells are able to self-renew having regeneration potential, and may be effective for the treatment of ovarian failure and consequently infertility. Brazilian Society of Assisted Reproduction 2020 /pmc/articles/PMC6993165/ /pubmed/31689043 http://dx.doi.org/10.5935/1518-0557.20190057 Text en http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Terraciano, Paula Barros
Garcez, Tuane Alves
Berger, Markus
Durli, Isabel
Kuhl, Cristiana Palma
Batista, Vitória de Oliveira
Schneider, Raquel de Almeida
Festa, Jaquelline
Pilar, Emily
Goulart, Marcela
Ferreira, Charles
Passos, Eduardo Pandolfi
Lima, Elizabeth Cirne
Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title_full Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title_fullStr Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title_full_unstemmed Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title_short Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice
title_sort ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in cf-1 mice
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6993165/
https://www.ncbi.nlm.nih.gov/pubmed/31689043
http://dx.doi.org/10.5935/1518-0557.20190057
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