Produção in vitro de embriões bovinos com cisteamina
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/10134 |
Resumo: | Advanced reproductive techniques use the 3 phase of in vitro production for bovine embryos, i.e. in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) as support for a variety of studies. This study assessed the effect of cysteamine, a thiol component, on the embryonic development during maturation, fertilization and in vitro culture. Bos taurus indicus Cumulus-oocyte complexes (COC) obtained from cow ovaries collected at a slaughterhouse were randomly distributed in four groups: controlgroup (without cysteamine, n=488), cysteamine in maturation (n=487), cysteamine in fertilization (n=489) and cysteamine in the culture medium (n=493). All COC were matured for 24h in TCM-199+0,01IUrFSH/mL+0,05mgLH/mL+10% fetal calf serum (FCS). In the cysteamine-maturation group, the TCM-199 medium was supplemented with 150μM cysteamine. The COC were matured at 38.5ºC with 5% of CO2 in air and humid atmosphere. The IVF (D0=fertilization day) was performed for 18-22h in Talp-Fert medium with heparin and PHE under the same conditions as IVM. The medium of the cysteamine-fertilization group was supplemented with 150μM cysteamine. For sperm capacitation, the semen was thawed, selected by percoll gradient and insemination was done with 2x106 spermatozoa/mL. Presumed zygotes from all groups were cultured in SOF + 5% FCS. In the cysteamine culture group the SOF medium was supplemented with 150μM cysteamine. Culture was maintained at 5%CO2, 5%O2, 90%N2 and saturated humidity for 8 days. Cleavage rates were 86, 89, 88 and 90 respectively, for control, maturation, fertilization and culture groups. Percentage of blastocyst on D7 were 29, 31, 38 e 35 and blastocyst hatched on D9 were 21, 23, 27 and 29, respectively. This study showed that adding cysteamine to the fertilization or culture medium improve the blastocyst production. Commercial OPU/IVP programs maybe benefit from the use of cysteamine. |
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Produção in vitro de embriões bovinos com cisteaminaIn vitro production of bovine embryos with cysteamineAntioxidantesPIVBovinosCisteaminaBos taurus indicusAntioxidantsIVPBovineCysteamineBos taurus indicusCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAAdvanced reproductive techniques use the 3 phase of in vitro production for bovine embryos, i.e. in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) as support for a variety of studies. This study assessed the effect of cysteamine, a thiol component, on the embryonic development during maturation, fertilization and in vitro culture. Bos taurus indicus Cumulus-oocyte complexes (COC) obtained from cow ovaries collected at a slaughterhouse were randomly distributed in four groups: controlgroup (without cysteamine, n=488), cysteamine in maturation (n=487), cysteamine in fertilization (n=489) and cysteamine in the culture medium (n=493). All COC were matured for 24h in TCM-199+0,01IUrFSH/mL+0,05mgLH/mL+10% fetal calf serum (FCS). In the cysteamine-maturation group, the TCM-199 medium was supplemented with 150μM cysteamine. The COC were matured at 38.5ºC with 5% of CO2 in air and humid atmosphere. The IVF (D0=fertilization day) was performed for 18-22h in Talp-Fert medium with heparin and PHE under the same conditions as IVM. The medium of the cysteamine-fertilization group was supplemented with 150μM cysteamine. For sperm capacitation, the semen was thawed, selected by percoll gradient and insemination was done with 2x106 spermatozoa/mL. Presumed zygotes from all groups were cultured in SOF + 5% FCS. In the cysteamine culture group the SOF medium was supplemented with 150μM cysteamine. Culture was maintained at 5%CO2, 5%O2, 90%N2 and saturated humidity for 8 days. Cleavage rates were 86, 89, 88 and 90 respectively, for control, maturation, fertilization and culture groups. Percentage of blastocyst on D7 were 29, 31, 38 e 35 and blastocyst hatched on D9 were 21, 23, 27 and 29, respectively. This study showed that adding cysteamine to the fertilization or culture medium improve the blastocyst production. Commercial OPU/IVP programs maybe benefit from the use of cysteamine.As biotécnicas reprodutivas atuais utilizam como suporte para outros estudos a produção in vitro (PIV) de embriões bovinos, conduzida em três etapas igualmente importantes: maturação in vitro (MIV), fecundação in vitro (FIV) e cultivo in vitro (CIV). Nesta pesquisa avaliou-se o efeito da cisteamina, um componente thiol, nos meios de maturação, fecundação ou de cultivo sobre o desenvolvimento embrionário. Complexos cumulusoócitos (CCO=oócitos) coletados de ovários de vacas Bos taurus indicus provenientes de frigorífico foram selecionados e distribuídos aleatoriamente em quatro grupos: Grupo Controle (sem cisteamina, n=488), Cisteamina na maturação (n=487), Cisteamina na fecundação (n=489) e Cisteamina no cultivo (n=493). Os oócitos dos quatro grupos foram maturados por 24h em TCM-199 modificado acrescido de 0,01UI/mL rFSHh + 0,05mg/mL de LH e 10% de SFB. No grupo cisteamina-maturação, o meio TCM-199 foi suplementado com 150μM de cisteamina. Os oócitos foram maturados a 38,5ºC em estufa com 5%CO2 em ar e umidade saturada. A FIV (D0=dia da fecendação) foi realizada por 18-22 horas em meio Fert-Talp acrescido de heparina e PHE. Este meio utilizado no grupo cisteamina-fecundação foi adicionado de 150μM de cisteamina. Para capacitação, o sêmen foi descongelado e processado através do Gradiente de Percoll e a inseminação foi realizada com 2x106 espermatozóides/mL, nas mesmas condições atmosféricas da MIV. Os prováveis zigotos de todos grupos experimentais foram cultivados em meio SOF suplementado com 5% SFB. No grupo cisteamina-cultivo o meio SOF foi acrescido de 150μM de cisteamina. A atmosfera gasosa nos 8 dias de cultivo foi de 5% de CO2, 5% de O2 e 90% de N2 e umidade saturada. A clivagem foi de 86, 89, 88 e 90%, obtendo-se 29, 31, 38 e 35% de blastocistos em D7 e 21, 23, 27 e 29% de eclosão para os grupos controle, maturação, fecundação e cultivo, respectivamente. Em meio de fecundação (Fert-Talp) ou em meio de cultivo (SOFaaci) a cisteamina promove maior produção embrionária. O uso da cisteamina nos programas comerciais de OPU/PIV poderá contribuir para melhor eficiência da produção embrionária.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaRubin, Mara Iolanda Batistellahttp://lattes.cnpq.br/5140943832465284Schossler, Deila Rosely Carneirohttp://lattes.cnpq.br/2268564401345390Bernardi, Mari Lourdeshttp://lattes.cnpq.br/1048010005537828Silva, Daniela Scherer da2017-06-012017-06-012006-02-21info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfSILVA, Daniela Scherer da. In vitro production of bovine embryos with cysteamine. 2006. 43 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2006.http://repositorio.ufsm.br/handle/1/10134porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2023-02-03T15:11:33Zoai:repositorio.ufsm.br:1/10134Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2023-02-03T15:11:33Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Produção in vitro de embriões bovinos com cisteamina In vitro production of bovine embryos with cysteamine |
title |
Produção in vitro de embriões bovinos com cisteamina |
spellingShingle |
Produção in vitro de embriões bovinos com cisteamina Silva, Daniela Scherer da Antioxidantes PIV Bovinos Cisteamina Bos taurus indicus Antioxidants IVP Bovine Cysteamine Bos taurus indicus CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Produção in vitro de embriões bovinos com cisteamina |
title_full |
Produção in vitro de embriões bovinos com cisteamina |
title_fullStr |
Produção in vitro de embriões bovinos com cisteamina |
title_full_unstemmed |
Produção in vitro de embriões bovinos com cisteamina |
title_sort |
Produção in vitro de embriões bovinos com cisteamina |
author |
Silva, Daniela Scherer da |
author_facet |
Silva, Daniela Scherer da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Rubin, Mara Iolanda Batistella http://lattes.cnpq.br/5140943832465284 Schossler, Deila Rosely Carneiro http://lattes.cnpq.br/2268564401345390 Bernardi, Mari Lourdes http://lattes.cnpq.br/1048010005537828 |
dc.contributor.author.fl_str_mv |
Silva, Daniela Scherer da |
dc.subject.por.fl_str_mv |
Antioxidantes PIV Bovinos Cisteamina Bos taurus indicus Antioxidants IVP Bovine Cysteamine Bos taurus indicus CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
topic |
Antioxidantes PIV Bovinos Cisteamina Bos taurus indicus Antioxidants IVP Bovine Cysteamine Bos taurus indicus CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
Advanced reproductive techniques use the 3 phase of in vitro production for bovine embryos, i.e. in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) as support for a variety of studies. This study assessed the effect of cysteamine, a thiol component, on the embryonic development during maturation, fertilization and in vitro culture. Bos taurus indicus Cumulus-oocyte complexes (COC) obtained from cow ovaries collected at a slaughterhouse were randomly distributed in four groups: controlgroup (without cysteamine, n=488), cysteamine in maturation (n=487), cysteamine in fertilization (n=489) and cysteamine in the culture medium (n=493). All COC were matured for 24h in TCM-199+0,01IUrFSH/mL+0,05mgLH/mL+10% fetal calf serum (FCS). In the cysteamine-maturation group, the TCM-199 medium was supplemented with 150μM cysteamine. The COC were matured at 38.5ºC with 5% of CO2 in air and humid atmosphere. The IVF (D0=fertilization day) was performed for 18-22h in Talp-Fert medium with heparin and PHE under the same conditions as IVM. The medium of the cysteamine-fertilization group was supplemented with 150μM cysteamine. For sperm capacitation, the semen was thawed, selected by percoll gradient and insemination was done with 2x106 spermatozoa/mL. Presumed zygotes from all groups were cultured in SOF + 5% FCS. In the cysteamine culture group the SOF medium was supplemented with 150μM cysteamine. Culture was maintained at 5%CO2, 5%O2, 90%N2 and saturated humidity for 8 days. Cleavage rates were 86, 89, 88 and 90 respectively, for control, maturation, fertilization and culture groups. Percentage of blastocyst on D7 were 29, 31, 38 e 35 and blastocyst hatched on D9 were 21, 23, 27 and 29, respectively. This study showed that adding cysteamine to the fertilization or culture medium improve the blastocyst production. Commercial OPU/IVP programs maybe benefit from the use of cysteamine. |
publishDate |
2006 |
dc.date.none.fl_str_mv |
2006-02-21 2017-06-01 2017-06-01 |
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 |
SILVA, Daniela Scherer da. In vitro production of bovine embryos with cysteamine. 2006. 43 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2006. http://repositorio.ufsm.br/handle/1/10134 |
identifier_str_mv |
SILVA, Daniela Scherer da. In vitro production of bovine embryos with cysteamine. 2006. 43 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2006. |
url |
http://repositorio.ufsm.br/handle/1/10134 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1805922161005690880 |