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Title: Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue
Authors: Celestino, Juliana Jales de Hollanda
Santos, Regiane Rodrigues dos
Lopes, Cláudio Afonso Pinho
Martins, Fabrício Sousa
Matos, Maria Helena Tavares
Melo, Mônica Aline Parente
Báo, Sônia Nair
Rodrigues, Ana Paula Ribeiro
Silva, José Roberto Viana
Figueiredo, José Ricardo de
Assunto:: Folículos pré-antrais
Bovino
Congelamento - tecido ovariano
Issue Date: 2008
Publisher: Elsevier
Citation: CELESTINO, Juliana Jales de Hollanda, et al. Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue. Animal Reproduction Science, v. 108, p. 309-318, 2008. Disponível em:<http://www.sciencedirect.com/science/article/pii/S0378432007003041>. Acesso em: 09 fev. 2015.
Abstract: Bovine preantral follicles within ovarian fragments were exposed and cryopreserved in absence or presence of 1.5 M glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO), undergoing a previous cooling at 20 °C for 1 h (protocol 1) or at 4 °C for 24 h (protocol 2) in 0.9% saline solution. At the end of each treatment, preantral follicles were classified as non-viable/viable when they were stained/not stained with trypan blue, respectively. To confirm viability staining, ultra-structure of the follicles was evaluated by transmission electronic microscopy (TEM). Data were compared by Chi-square test (P < 0.05). The storage of the ovaries at 20 °C for 1 h (78%) and 4 °C for 24 h (80%) did not reduce significantly the percentage of viable preantral follicles when compared to the control (75%). Similar results were obtained when ovarian fragments, respectively, for protocols 1 and 2, were exposed to MEM (78 and 77%), 1.5 M EG (78 and 71%), as well as frozen in 1.5 M EG (74 and 77%). Percentages of viable follicles in control were similar to those observed after exposure (75%) and freezing (76%) in presence of 1.5 M DMSO only when protocol 1 was used. The increase of the concentration from 1.5 to 3.0 M, for all cryoprotectants, reduced significantly the percentage of viable preantral follicles after freezing. Ultra-structural analysis has confirmed trypan blue results, showing that not only basement membrane, but also organelles, were intact in viable preantral follicles. In conclusion, ovarian tissue cooling at 4 °C for 24 h before cryopreservation (protocol 2) does not affect the viability of bovine preantral follicles when 1.5 M EG is present in the cryopreservation medium.
Licença:: Este Manuscrito do Autor Aceito para Publicação (AAM) é protegido por direitos autorais e publicado pela Elsevier. Ele esta disponível neste Repositório, por acordo entre a Elsevier e a Universidade de Brasília. As alterações decorrentes do processo de publicação - como a edição, correção, formatação estrutural, e outros mecanismos de controle de qualidade - não estão refletidas nesta versão do texto. A versão definitiva do texto foi posteriormente publicado em [Animal Reproduction Science, Volume 108, Número 3-4, Novembro de 2008, Páginas 309–318 doi:10.1016/j.anireprosci.2007.08.016]. Você pode baixar, copiar e utilizar de outra forma o AAM para fins não comerciais , desde que sua licença seja limitada pelas seguintes restrições: (1) Você pode usar este AAM para fins não comerciais apenas sob os termos da licença CC- BY- NC-ND. (2) A integridade do trabalho e identificação do autor, detentor dos direitos autorais e editor deve ser preservado em qualquer cópia. (3) Tem de atribuir este AAM no seguinte formato: [acordo na linguagem atribuída, incluindo o link para CC BY-NC-ND licença Digital + DOI do artigo publicado na revista Elsevier ScienceDirect ® da plataforma].
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