Aspectos histológicos do ovário de coelhas após criopreservação
Autor(a) principal: | |
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Data de Publicação: | 2005 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://dx.doi.org/10.1590/S0100-72032005001100002 http://repositorio.unifesp.br/handle/11600/2747 |
Resumo: | PURPOSE: to evaluate follicular preservation and histologic characteristics of the cryopreserved ovarian tissue and to compare with the fresh one, in rabbits. METHODS: ten adult female white rabbits were submitted to right oophorectomy. The dried ovary was dissected and the cortex was maintained with approximately 1.5 millimeter thickness. The tissue was fractionated into small sections, some reserved for control histologic study and others destined for cryopreservation. Six weeks later the ovarian tissue was thawed and evaluated histologically. After histologic processing, the control and the experimental samples were stained with hematoxylin and eosin and treated immunohistochemically by the PCNA technique for evaluation of DNA preservation. Histologic alterations present in the fresh and in the cryopreserved tissues were identified, and cryopreserved tissue viability was evaluated. RESULTS: in the cryopreserved tissue only primordial follicles persisted. Reversible alterations were identified: cytoplasmatic vacuolation (p=0,039), stromal lysis (p=0.648) and oocytes with irregular contours (p=0.007). Irreversible alterations: (hyalin degeneration and pyknosis) were found, but not at significant levels (p=0.210). The immunohistochemical analysis showed PCNA staining of follicles at different stages of development in the fresh tissue and primordial follicles in the cryopreserved tissue, indicating the presence of active DNA in both tissues. CONCLUSION: in the cryopreserved ovarian tissue the following were observed: survival of only primordial follicles; significant reversible histologic alterations (cytoplasmic vacuolation, stromal lysis and oocytes with irregular contours); irreversible alterations (hyalin degeneration and pyknosis), and PCNA staining of all follicles. |
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Aspectos histológicos do ovário de coelhas após criopreservaçãoHistological aspects of rabbit ovarian tissue after cryopreservationCryopreservationOvaryPloriferating cell nuclear antigemRabbitsCriopreservaçãoOvárioAntigeno nuclear de células em proliferaçãoCoelhosPURPOSE: to evaluate follicular preservation and histologic characteristics of the cryopreserved ovarian tissue and to compare with the fresh one, in rabbits. METHODS: ten adult female white rabbits were submitted to right oophorectomy. The dried ovary was dissected and the cortex was maintained with approximately 1.5 millimeter thickness. The tissue was fractionated into small sections, some reserved for control histologic study and others destined for cryopreservation. Six weeks later the ovarian tissue was thawed and evaluated histologically. After histologic processing, the control and the experimental samples were stained with hematoxylin and eosin and treated immunohistochemically by the PCNA technique for evaluation of DNA preservation. Histologic alterations present in the fresh and in the cryopreserved tissues were identified, and cryopreserved tissue viability was evaluated. RESULTS: in the cryopreserved tissue only primordial follicles persisted. Reversible alterations were identified: cytoplasmatic vacuolation (p=0,039), stromal lysis (p=0.648) and oocytes with irregular contours (p=0.007). Irreversible alterations: (hyalin degeneration and pyknosis) were found, but not at significant levels (p=0.210). The immunohistochemical analysis showed PCNA staining of follicles at different stages of development in the fresh tissue and primordial follicles in the cryopreserved tissue, indicating the presence of active DNA in both tissues. CONCLUSION: in the cryopreserved ovarian tissue the following were observed: survival of only primordial follicles; significant reversible histologic alterations (cytoplasmic vacuolation, stromal lysis and oocytes with irregular contours); irreversible alterations (hyalin degeneration and pyknosis), and PCNA staining of all follicles.OBJETIVO: avaliar a preservação folicular e as características celulares do tecido ovariano criopreservado, em coelhas. MÉTODOS: fez-se, sob anestesia, a ooforectomia direita de dez coelhas brancas, adultas. Dissecou-se o ovário mantendo-se o córtex com espessura de 1,5 milímetros. Fragmentou-se o tecido em pequenas secções, algumas para o estudo histológico de controle e outras destinadas à criopreservação pelo protocolo de congelamento lento. Passadas seis semanas efetuou-se o descongelamento e fez-se a avaliação histológica. As amostras do controle e do experimento, após processamento, foram coradas pela hematoxilina-eosina para identificação dos aspectos histológicos e submetidas à técnica imuno-histoquímica utilizando-se o PCNA (proliferating cell nuclear antigen) para a avaliação da viabilidade celular. Utilizaram-se os testes não paramétricos comparação entre duas proporções e Mann-Whitney e o teste paramétrico t de Student. RESULTADOS: observou-se que no tecido criopreservado só persistiram oócitos primordiais. Entre as alterações reversíveis identificaram-se: vacuolização citoplasmática em todas as amostras (p=0,039), lise estromal em 50% (p=0,648) e oócitos com contornos irregulares em 80% (p=0,007). Encontraram-se alterações irreversíveis como degeneração hialina e picnose em 30% das amostras (p=0,210). A análise imuno-histoquímica demonstrou os folículos, em diferentes estágios de desenvolvimento, no tecido não congelado e folículos primordiais no tecido criopreservado com positividade para o PCNA, indicando a presença de DNA ativo. CONCLUSÃO: no tecido ovariano criopreservado sobrevivem apenas os folículos primordiais; existem alterações histológicas reversíveis (vacuolização citoplasmática, lise estromal, fragmentação das células da granulosa e oócitos com contornos irregulares); alterações irreversíveis, em níveis não significantes (degeneração hialina e picnose) e presença de PCNA positivo em todos os folículos.PUCPR Instituto de FertilidadeUniversidade Federal de São Paulo (UNIFESP) EPMUniversidade Federal do Paraná Departamento de CirurgiaColégio Brasileiro de CirurgiõesMaterbaby Reprodução Humana e Genéticae Genética Universidade Estadual de MaringáFaculdade de Medicina Evangélica do Paraná Instituto de Pesquisas MédicasUniversidade de MieUNIFESP, EPMSciELOFederação Brasileira das Sociedades de Ginecologia e ObstetríciaPUCPR Instituto de FertilidadeUniversidade Federal de São Paulo (UNIFESP)Universidade Federal do Paraná Departamento de CirurgiaColégio Brasileiro de CirurgiõesMaterbaby Reprodução Humana e Genéticae Genética Universidade Estadual de MaringáFaculdade de Medicina Evangélica do Paraná Instituto de Pesquisas MédicasUniversidade de MieThomaz, Beatriz Angélica CharlotteBiondo-simões, Maria De Lourdes Pessole [UNIFESP]Almodin, Carlos Gilberto [UNIFESP]Minguetti-camara, Vânia CibeleCeschin, Álvaro PigatoIoshii, Sergio Ossamu2015-06-14T13:31:47Z2015-06-14T13:31:47Z2005-11-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion642-649application/pdfhttp://dx.doi.org/10.1590/S0100-72032005001100002Revista Brasileira de Ginecologia e Obstetrícia. Federação Brasileira das Sociedades de Ginecologia e Obstetrícia, v. 27, n. 11, p. 642-649, 2005.10.1590/S0100-72032005001100002S0100-72032005001100002.pdf0100-7203S0100-72032005001100002http://repositorio.unifesp.br/handle/11600/2747porRevista Brasileira de Ginecologia e Obstetríciainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-04T14:20:15Zoai:repositorio.unifesp.br/:11600/2747Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-04T14:20:15Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.none.fl_str_mv |
Aspectos histológicos do ovário de coelhas após criopreservação Histological aspects of rabbit ovarian tissue after cryopreservation |
title |
Aspectos histológicos do ovário de coelhas após criopreservação |
spellingShingle |
Aspectos histológicos do ovário de coelhas após criopreservação Thomaz, Beatriz Angélica Charlotte Cryopreservation Ovary Ploriferating cell nuclear antigem Rabbits Criopreservação Ovário Antigeno nuclear de células em proliferação Coelhos |
title_short |
Aspectos histológicos do ovário de coelhas após criopreservação |
title_full |
Aspectos histológicos do ovário de coelhas após criopreservação |
title_fullStr |
Aspectos histológicos do ovário de coelhas após criopreservação |
title_full_unstemmed |
Aspectos histológicos do ovário de coelhas após criopreservação |
title_sort |
Aspectos histológicos do ovário de coelhas após criopreservação |
author |
Thomaz, Beatriz Angélica Charlotte |
author_facet |
Thomaz, Beatriz Angélica Charlotte Biondo-simões, Maria De Lourdes Pessole [UNIFESP] Almodin, Carlos Gilberto [UNIFESP] Minguetti-camara, Vânia Cibele Ceschin, Álvaro Pigato Ioshii, Sergio Ossamu |
author_role |
author |
author2 |
Biondo-simões, Maria De Lourdes Pessole [UNIFESP] Almodin, Carlos Gilberto [UNIFESP] Minguetti-camara, Vânia Cibele Ceschin, Álvaro Pigato Ioshii, Sergio Ossamu |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
PUCPR Instituto de Fertilidade Universidade Federal de São Paulo (UNIFESP) Universidade Federal do Paraná Departamento de Cirurgia Colégio Brasileiro de Cirurgiões Materbaby Reprodução Humana e Genética e Genética Universidade Estadual de Maringá Faculdade de Medicina Evangélica do Paraná Instituto de Pesquisas Médicas Universidade de Mie |
dc.contributor.author.fl_str_mv |
Thomaz, Beatriz Angélica Charlotte Biondo-simões, Maria De Lourdes Pessole [UNIFESP] Almodin, Carlos Gilberto [UNIFESP] Minguetti-camara, Vânia Cibele Ceschin, Álvaro Pigato Ioshii, Sergio Ossamu |
dc.subject.por.fl_str_mv |
Cryopreservation Ovary Ploriferating cell nuclear antigem Rabbits Criopreservação Ovário Antigeno nuclear de células em proliferação Coelhos |
topic |
Cryopreservation Ovary Ploriferating cell nuclear antigem Rabbits Criopreservação Ovário Antigeno nuclear de células em proliferação Coelhos |
description |
PURPOSE: to evaluate follicular preservation and histologic characteristics of the cryopreserved ovarian tissue and to compare with the fresh one, in rabbits. METHODS: ten adult female white rabbits were submitted to right oophorectomy. The dried ovary was dissected and the cortex was maintained with approximately 1.5 millimeter thickness. The tissue was fractionated into small sections, some reserved for control histologic study and others destined for cryopreservation. Six weeks later the ovarian tissue was thawed and evaluated histologically. After histologic processing, the control and the experimental samples were stained with hematoxylin and eosin and treated immunohistochemically by the PCNA technique for evaluation of DNA preservation. Histologic alterations present in the fresh and in the cryopreserved tissues were identified, and cryopreserved tissue viability was evaluated. RESULTS: in the cryopreserved tissue only primordial follicles persisted. Reversible alterations were identified: cytoplasmatic vacuolation (p=0,039), stromal lysis (p=0.648) and oocytes with irregular contours (p=0.007). Irreversible alterations: (hyalin degeneration and pyknosis) were found, but not at significant levels (p=0.210). The immunohistochemical analysis showed PCNA staining of follicles at different stages of development in the fresh tissue and primordial follicles in the cryopreserved tissue, indicating the presence of active DNA in both tissues. CONCLUSION: in the cryopreserved ovarian tissue the following were observed: survival of only primordial follicles; significant reversible histologic alterations (cytoplasmic vacuolation, stromal lysis and oocytes with irregular contours); irreversible alterations (hyalin degeneration and pyknosis), and PCNA staining of all follicles. |
publishDate |
2005 |
dc.date.none.fl_str_mv |
2005-11-01 2015-06-14T13:31:47Z 2015-06-14T13:31:47Z |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1590/S0100-72032005001100002 Revista Brasileira de Ginecologia e Obstetrícia. Federação Brasileira das Sociedades de Ginecologia e Obstetrícia, v. 27, n. 11, p. 642-649, 2005. 10.1590/S0100-72032005001100002 S0100-72032005001100002.pdf 0100-7203 S0100-72032005001100002 http://repositorio.unifesp.br/handle/11600/2747 |
url |
http://dx.doi.org/10.1590/S0100-72032005001100002 http://repositorio.unifesp.br/handle/11600/2747 |
identifier_str_mv |
Revista Brasileira de Ginecologia e Obstetrícia. Federação Brasileira das Sociedades de Ginecologia e Obstetrícia, v. 27, n. 11, p. 642-649, 2005. 10.1590/S0100-72032005001100002 S0100-72032005001100002.pdf 0100-7203 S0100-72032005001100002 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
Revista Brasileira de Ginecologia e Obstetrícia |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
642-649 application/pdf |
dc.publisher.none.fl_str_mv |
Federação Brasileira das Sociedades de Ginecologia e Obstetrícia |
publisher.none.fl_str_mv |
Federação Brasileira das Sociedades de Ginecologia e Obstetrícia |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
biblioteca.csp@unifesp.br |
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1814268315343781888 |