Vitrificação de oócitos bovinos imaturos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2002 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/26734 |
Resumo: | The development of reproductive biotechnology has been originating new challenges in many areas. In the cryobiology, the current challenge is to obtain an appropriate methodology for bovine oocyte cryopreservation, especially in the immature stage. This study was aimed to the evaluate the in vitro and in vivo development of embryos derived from immature vitrified bovine oocytes, with OPS technology. In addition was evaluate the effect of the pre-treatment with cytochalasin D (CD), and the viability of a cryoprotectant solution composed by ethylene glycol (EG), dimethyl sulfoxide (Me2SO) and sucrose (SUC), in the vitrification of immature bovine oocytes. The oocytes obtained of slaughterhouse bovine ovaries were randomly allocated in three groups: a control group (n=442), a vitrified group (Vitri n=356) and a group treated with 20μg/ml of CD during 10 to 20 minutes and vitrified (CDVitri n=355). For the vitrification procedure, the oocytes were exposed for 30 seconds to an VS50 solution (10% EG + 10% DMSO), followed by 25 seconds exposure to a VS100 solution (20% EG + 20% DMSO + 0.5M of SUC), loaded in groups of 4 to 5 in OPS and plunged in liquid nitrogen. The warming was performed in two steps of 5 minutes, with sucrose gradients (0.26 and 0.16M) solutions. After that, all groups were submitted to the same in vitro maturation, fertilization and culture procedures. The in vitro viability was evaluated by the cleavage and blastocysts rates, and the in vivo viability was evaluated by the transfer of blastocysts derived from vitrified immature oocytes (1 or 2 for each recipient). No differences (P>0.05) were observed in the cleavage and blastocysts rates among the CDVitri (46.4% and 3.5%) and Vitri (49.0% and 6.1%) groups, respectively, and both were lower (p<0.05) than control group (85.1% and 45.9%). A female calf was obtained from CDVitri group (two recipients) and a female and a male calves birth from the Vitri group (five recipients), were obtained. These data indicate that the cytochalasin D pre-treatment do not improve viability of embryos derived from vitrified immature bovine oocytes. The OPS technology with a cryoprotectant solution composed by 20% EG + 20% Me2SO and 0.5M SUC is effective for vitrification of immature oocytes. The embryos obtained from immature oocytes vitrified with this technology are fully competent to develop healthy offspring, even when submitted to a new vitrification procedure at blastocyst stage. |
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2022-11-01T18:22:00Z2022-11-01T18:22:00Z2002-02-22http://repositorio.ufsm.br/handle/1/26734The development of reproductive biotechnology has been originating new challenges in many areas. In the cryobiology, the current challenge is to obtain an appropriate methodology for bovine oocyte cryopreservation, especially in the immature stage. This study was aimed to the evaluate the in vitro and in vivo development of embryos derived from immature vitrified bovine oocytes, with OPS technology. In addition was evaluate the effect of the pre-treatment with cytochalasin D (CD), and the viability of a cryoprotectant solution composed by ethylene glycol (EG), dimethyl sulfoxide (Me2SO) and sucrose (SUC), in the vitrification of immature bovine oocytes. The oocytes obtained of slaughterhouse bovine ovaries were randomly allocated in three groups: a control group (n=442), a vitrified group (Vitri n=356) and a group treated with 20μg/ml of CD during 10 to 20 minutes and vitrified (CDVitri n=355). For the vitrification procedure, the oocytes were exposed for 30 seconds to an VS50 solution (10% EG + 10% DMSO), followed by 25 seconds exposure to a VS100 solution (20% EG + 20% DMSO + 0.5M of SUC), loaded in groups of 4 to 5 in OPS and plunged in liquid nitrogen. The warming was performed in two steps of 5 minutes, with sucrose gradients (0.26 and 0.16M) solutions. After that, all groups were submitted to the same in vitro maturation, fertilization and culture procedures. The in vitro viability was evaluated by the cleavage and blastocysts rates, and the in vivo viability was evaluated by the transfer of blastocysts derived from vitrified immature oocytes (1 or 2 for each recipient). No differences (P>0.05) were observed in the cleavage and blastocysts rates among the CDVitri (46.4% and 3.5%) and Vitri (49.0% and 6.1%) groups, respectively, and both were lower (p<0.05) than control group (85.1% and 45.9%). A female calf was obtained from CDVitri group (two recipients) and a female and a male calves birth from the Vitri group (five recipients), were obtained. These data indicate that the cytochalasin D pre-treatment do not improve viability of embryos derived from vitrified immature bovine oocytes. The OPS technology with a cryoprotectant solution composed by 20% EG + 20% Me2SO and 0.5M SUC is effective for vitrification of immature oocytes. The embryos obtained from immature oocytes vitrified with this technology are fully competent to develop healthy offspring, even when submitted to a new vitrification procedure at blastocyst stage.A constante evolução das biotécnicas da reprodução tem originado novos desafios em diferentes áreas. Na criobiologia, o atual desafio é a obtenção de uma adequada metodologia para a criopreservação de oócitos bovinos, especialmente no estágio imaturo. Com este propósito, foi conduzido o presente estudo que tem como objetivos avaliar o desenvolvimento in vitro e in vivo de embriões derivados de oócitos imaturos vitrificados com a tecnologia OPS, além de avaliar o efeito do pré-tratamento com citocalasina D (CD) e a eficácia da solução crioprotetora composta por 20% de etileno glicol (EG) + 20% de dimetil sulfóxido (Me2SO) + 0.5 M de sacarose (SAC). Oócitos obtidos de ovários de matadouro foram aleatoriamente distribuídos em três grupos: controle (n=442), vitrificado (Vitri n=356) e tratado com 20μg/ml de CD durante 10 a 20 minutos e vitrificado (CDVitri n=355). Para a vitrificação, os oócitos foram expostos durante 30 segundos a solução SV50 (10% EG + 10% Me2SO), seguido de 25 segundos na solução SV100 (20% EG+ 20% Me2SO + 0,5M SAC), sendo envasados em grupos de 4 a 5 em cada OPS e mergulhados em nitrogênio líquido. O aquecimento foi realizado em dois passos de 5 minutos, com gradientes de sacarose (0,26 e 0,16M). Após isso, todos os grupos foram submetidos simultaneamente aos mesmos processos de maturação, fecundação e cultivo in vitro. A viabilidade in vitro foi avaliada através das taxas de clivagem e blastocistos. Adicionalmente, a viabilidade in vivo foi avaliada pela transferência dos embriões (1 ou 2 por receptora), derivados de oócitos imaturos vitrificados, submetidos ou não a nova vitrificação no estágio de blastocisto. Não foram observadas diferenças estatísticas nas taxas de clivagem e blastocistos entre os grupos CDVitri (46,4% e 3,5%) e Vitri (49,0% e 6,1%) respectivamente, sendo ambos inferiores (P<0,05) ao grupo controle (85,1% e 45,9%). Na avaliação in vivo, obteve-se o nascimento de uma fêmea e um macho do grupo Vitri (cinco receptoras) e uma fêmea do grupo CDVitri (duas receptoras). Esses resultados permitem concluir que o pré-tratamento com citocalasina D não melhora a viabilidade embrionária de oócitos bovinos imaturos vitrificados. Entretanto, a solução de vitrificação utilizada foi eficiente para criopreservação de oócitos imaturos, com a tecnologia OPS. Os embriões derivados de oócitos imaturos vitrificados possibilitam a obtenção do nascimento de produtos normais e saudáveis, mesmo quando submetidos a nova vitrificação no estágio de blastocisto expandido.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqporUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Medicina VeterináriaUFSMBrasilMedicina VeterináriaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessBovinoOócito imaturoVitrificaçãoOPSCitocalasina DBlastocistosBovineImmature oocyteVitrificationCytochalasin DBlastocystsCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAVitrificação de oócitos bovinos imaturosImmature bovine oocytes vitrificationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisRubin, Mara Iolanda Batistellahttp://lattes.cnpq.br/5140943832465284Rheingantz, Maria Gabriela TavaresAzambuja, Ricardo Marques dehttp://lattes.cnpq.br/6064420287868085Vieira, Arnaldo Diniz50050000000760060060060060002a1a2f5-cf70-4065-ae1f-c6bba79e884891f77f71-7194-431c-8825-075f4c23fd028c5af777-f5ea-4ebc-8047-8cf15686712aeeae7759-3405-4823-bea1-2e6628e89860reponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALDIS_PPGMV_2002_VIEIRA _ARNALDO.pdfDIS_PPGMV_2002_VIEIRA _ARNALDO.pdfDissertação de mestradoapplication/pdf712469http://repositorio.ufsm.br/bitstream/1/26734/1/DIS_PPGMV_2002_VIEIRA%20_ARNALDO.pdf053330a4aba1a63ba3cf21492397513cMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81956http://repositorio.ufsm.br/bitstream/1/26734/3/license.txt2f0571ecee68693bd5cd3f17c1e075dfMD53CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.ufsm.br/bitstream/1/26734/2/license_rdf4460e5956bc1d1639be9ae6146a50347MD521/267342022-11-01 15:22:00.681oai:repositorio.ufsm.br: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ório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-11-01T18:22Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Vitrificação de oócitos bovinos imaturos |
dc.title.alternative.eng.fl_str_mv |
Immature bovine oocytes vitrification |
title |
Vitrificação de oócitos bovinos imaturos |
spellingShingle |
Vitrificação de oócitos bovinos imaturos Vieira, Arnaldo Diniz Bovino Oócito imaturo Vitrificação OPS Citocalasina D Blastocistos Bovine Immature oocyte Vitrification Cytochalasin D Blastocysts CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Vitrificação de oócitos bovinos imaturos |
title_full |
Vitrificação de oócitos bovinos imaturos |
title_fullStr |
Vitrificação de oócitos bovinos imaturos |
title_full_unstemmed |
Vitrificação de oócitos bovinos imaturos |
title_sort |
Vitrificação de oócitos bovinos imaturos |
author |
Vieira, Arnaldo Diniz |
author_facet |
Vieira, Arnaldo Diniz |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Rubin, Mara Iolanda Batistella |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5140943832465284 |
dc.contributor.referee1.fl_str_mv |
Rheingantz, Maria Gabriela Tavares |
dc.contributor.referee2.fl_str_mv |
Azambuja, Ricardo Marques de |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6064420287868085 |
dc.contributor.author.fl_str_mv |
Vieira, Arnaldo Diniz |
contributor_str_mv |
Rubin, Mara Iolanda Batistella Rheingantz, Maria Gabriela Tavares Azambuja, Ricardo Marques de |
dc.subject.por.fl_str_mv |
Bovino Oócito imaturo Vitrificação OPS Citocalasina D Blastocistos |
topic |
Bovino Oócito imaturo Vitrificação OPS Citocalasina D Blastocistos Bovine Immature oocyte Vitrification Cytochalasin D Blastocysts CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.eng.fl_str_mv |
Bovine Immature oocyte Vitrification Cytochalasin D Blastocysts |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The development of reproductive biotechnology has been originating new challenges in many areas. In the cryobiology, the current challenge is to obtain an appropriate methodology for bovine oocyte cryopreservation, especially in the immature stage. This study was aimed to the evaluate the in vitro and in vivo development of embryos derived from immature vitrified bovine oocytes, with OPS technology. In addition was evaluate the effect of the pre-treatment with cytochalasin D (CD), and the viability of a cryoprotectant solution composed by ethylene glycol (EG), dimethyl sulfoxide (Me2SO) and sucrose (SUC), in the vitrification of immature bovine oocytes. The oocytes obtained of slaughterhouse bovine ovaries were randomly allocated in three groups: a control group (n=442), a vitrified group (Vitri n=356) and a group treated with 20μg/ml of CD during 10 to 20 minutes and vitrified (CDVitri n=355). For the vitrification procedure, the oocytes were exposed for 30 seconds to an VS50 solution (10% EG + 10% DMSO), followed by 25 seconds exposure to a VS100 solution (20% EG + 20% DMSO + 0.5M of SUC), loaded in groups of 4 to 5 in OPS and plunged in liquid nitrogen. The warming was performed in two steps of 5 minutes, with sucrose gradients (0.26 and 0.16M) solutions. After that, all groups were submitted to the same in vitro maturation, fertilization and culture procedures. The in vitro viability was evaluated by the cleavage and blastocysts rates, and the in vivo viability was evaluated by the transfer of blastocysts derived from vitrified immature oocytes (1 or 2 for each recipient). No differences (P>0.05) were observed in the cleavage and blastocysts rates among the CDVitri (46.4% and 3.5%) and Vitri (49.0% and 6.1%) groups, respectively, and both were lower (p<0.05) than control group (85.1% and 45.9%). A female calf was obtained from CDVitri group (two recipients) and a female and a male calves birth from the Vitri group (five recipients), were obtained. These data indicate that the cytochalasin D pre-treatment do not improve viability of embryos derived from vitrified immature bovine oocytes. The OPS technology with a cryoprotectant solution composed by 20% EG + 20% Me2SO and 0.5M SUC is effective for vitrification of immature oocytes. The embryos obtained from immature oocytes vitrified with this technology are fully competent to develop healthy offspring, even when submitted to a new vitrification procedure at blastocyst stage. |
publishDate |
2002 |
dc.date.issued.fl_str_mv |
2002-02-22 |
dc.date.accessioned.fl_str_mv |
2022-11-01T18:22:00Z |
dc.date.available.fl_str_mv |
2022-11-01T18:22:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/26734 |
url |
http://repositorio.ufsm.br/handle/1/26734 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500500000007 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 |
dc.relation.authority.fl_str_mv |
02a1a2f5-cf70-4065-ae1f-c6bba79e8848 91f77f71-7194-431c-8825-075f4c23fd02 8c5af777-f5ea-4ebc-8047-8cf15686712a eeae7759-3405-4823-bea1-2e6628e89860 |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Medicina Veterinária |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
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