Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/18853 http://doi.org/10.14393/ufu.di.2017.222 |
Resumo: | The aim of this study was to evaluate the development of secondary follicles in the in vitro culture on a three-dimensional system of nanoparticles (gold, iron oxide and poly-L-lysine) associated with magnetic levitation at three concentrations (50, 100 and 200 μL/mL) compared to 2D in vitro culture (control, traditional) for 16 days. Sixty bovine ovaries were collected and the micro dissection of the secondary preantral follicles (200 μm to 400 μm in diameter) was done manually from the fragments of the ovarian cortex. Thereafter, the isolated follicles were individually transferred to micro tubes containing culture medium. For 3D treatments, the nanoparticles were added according to the proposed concentrations. After 12 hours of incubation, the follicles were transferred to 24-well plates with culture medium and cultured for 16 days. At the end of the culture, the oocytes were in vitro matured for 22-24 hours. A total of 213 preantral follicles were selected for in vitro culture and randomly assigned to the treatments. The 200 μL/mL treatment showed a reduction (P < 0.05) in follicular viability only from D12 and at the end of the culture (D16), the highest proportion of viable follicles (P < 0.05) was observed in this group. The treatments with the highest concentrations of nanoparticles had the lowest rates of oocyte extrusion and higher proportion of antrum formation. In addition, within the slow-growing category, a superior percentage of follicles (P < 0.05) was observed in the 200 μL/mL group. The follicles of the control group were 6 times more likely (P < 0.05) of follicular degeneration and/or extrusion when compared to 200 μL/mL group. Additionally, the oocyte viability rate in in vitro maturation was higher (P < 0.05) in the treatment of 200 μL/mL. Furthermore, only the oocytes developed in the 3D culture reached the MI phase after the resumption of meiosis. Finally, it was concluded that the three-dimensional culture system with nanoparticles associated with magnetic levitation promoted the in vitro development of bovine secondary follicles and the concentration of 200 μL/mL obtained the highest proportion of viable follicles with the best rates of antrum formation and maintenance. |
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Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnéticaVeterináriaBovino - ReproduçãoFertilização in vitroNanopartículasFolículos pré-antraisCultivo tridimensionalLevitação magnéticaNanoparticlesMagnetic levitationPreantral folliclesThree-dimensional cultureCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAThe aim of this study was to evaluate the development of secondary follicles in the in vitro culture on a three-dimensional system of nanoparticles (gold, iron oxide and poly-L-lysine) associated with magnetic levitation at three concentrations (50, 100 and 200 μL/mL) compared to 2D in vitro culture (control, traditional) for 16 days. Sixty bovine ovaries were collected and the micro dissection of the secondary preantral follicles (200 μm to 400 μm in diameter) was done manually from the fragments of the ovarian cortex. Thereafter, the isolated follicles were individually transferred to micro tubes containing culture medium. For 3D treatments, the nanoparticles were added according to the proposed concentrations. After 12 hours of incubation, the follicles were transferred to 24-well plates with culture medium and cultured for 16 days. At the end of the culture, the oocytes were in vitro matured for 22-24 hours. A total of 213 preantral follicles were selected for in vitro culture and randomly assigned to the treatments. The 200 μL/mL treatment showed a reduction (P < 0.05) in follicular viability only from D12 and at the end of the culture (D16), the highest proportion of viable follicles (P < 0.05) was observed in this group. The treatments with the highest concentrations of nanoparticles had the lowest rates of oocyte extrusion and higher proportion of antrum formation. In addition, within the slow-growing category, a superior percentage of follicles (P < 0.05) was observed in the 200 μL/mL group. The follicles of the control group were 6 times more likely (P < 0.05) of follicular degeneration and/or extrusion when compared to 200 μL/mL group. Additionally, the oocyte viability rate in in vitro maturation was higher (P < 0.05) in the treatment of 200 μL/mL. Furthermore, only the oocytes developed in the 3D culture reached the MI phase after the resumption of meiosis. Finally, it was concluded that the three-dimensional culture system with nanoparticles associated with magnetic levitation promoted the in vitro development of bovine secondary follicles and the concentration of 200 μL/mL obtained the highest proportion of viable follicles with the best rates of antrum formation and maintenance.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorCNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo a Pesquisa do Estado de Minas GeraisDissertação (Mestrado)Objetivou-se avaliar o desenvolvimento dos folículos secundários de bovinos no cultivo in vitro em sistema 3D de nanopartículas (ouro, óxido de ferro e poli-L-lisina) associadas a levitação magnética com três concentrações (50, 100 e 200 μL/mL) comparado ao cultivo in vitro 2D (controle, cultivo tradicional em placa) durante 16 dias. Foram coletados 60 ovários bovinos e a microdissecção dos folículos pré-antrais secundários 200 μm a 400 μm de diâmetro foi feita a partir dos fragmentos do córtex ovariano de forma manual. Em seguida, os folículos isolados foram transferidos de forma individual para microtubos que continham meio de cultivo. Para os tratamentos de cultura em 3D, foram adicionadas as nanopartículas de acordo com as concentrações propostas. Após 12 horas de incubação, os folículos foram transferidos para placas de 24 poços com meio de cultivo e permaneceram novamente na estufa por 16 dias. Ao final do cultivo, os ovócitos foram maturados in vitro por 22-24 horas. Um total de 213 folículos pré-antrais foram selecionados para o cultivo in vitro e distribuídos aleatoriamente nos tratamentos. O tratamento 200 μL/mL apresentou redução (P < 0,05) na viabilidade folicular apenas a partir do D12 e ao final do cultivo (D16), a maior proporção de folículos viáveis (P < 0,05) foi observada neste grupo. Os tratamentos com as maiores concentrações de nanopartículas apresentaram as menores taxas de extrusão ovocitária e maior proporção de formação de antro. Além disso, dentro da categoria de crescimento lento, uma porcentagem de superior folículos (P < 0,05) foi observada no grupo 200 μL/mL. Os folículos do grupo controle apresentaram 6 vezes mais chances (P < 0,05) de degeneração e/ou extrusão folicular quando comparado ao 200 μL/mL. Adicionalmente, a taxa de viabilidade ovocitária na maturação in vitro foi superior (P < 0,05) no tratamento de 200 μL/mL apenas os ovócitos desenvolvidos no cultivo 3D alcançaram a fase de MI após a retomada da meiose. Finalmente, concluiu-se que o sistema de cultivo tridimensional com nanopartículas associadas a levitação magnética propiciou o desenvolvimento in vitro dos folículos secundários bovinos e a concentração de 200 μL/mL obteve a maior proporção de folículos viáveis com a melhores taxas de formação e manutenção do antro.Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Ciências VeterináriasAlves, Kele Amaralhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4297620J0Jacomini, Jose Octaviohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782265U4Vieira, Luis AlbertoBeletti, Marcelo Emíliohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703947D5Alves, Bênner Geraldohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4259761D1Antonino, Deize de Cássia2017-06-13T12:37:11Z2017-06-13T12:37:11Z2017-02-24info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfANTONINO, Deize de Cássia. Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética. 2017. 71 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Uberlândia, Uberlândia, 2017. DOI http://doi.org/10.14393/ufu.di.2017.222https://repositorio.ufu.br/handle/123456789/18853http://doi.org/10.14393/ufu.di.2017.222porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2021-09-20T19:13:26Zoai:repositorio.ufu.br:123456789/18853Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2021-09-20T19:13:26Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
title |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
spellingShingle |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética Antonino, Deize de Cássia Veterinária Bovino - Reprodução Fertilização in vitro Nanopartículas Folículos pré-antrais Cultivo tridimensional Levitação magnética Nanoparticles Magnetic levitation Preantral follicles Three-dimensional culture CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
title_full |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
title_fullStr |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
title_full_unstemmed |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
title_sort |
Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética |
author |
Antonino, Deize de Cássia |
author_facet |
Antonino, Deize de Cássia |
author_role |
author |
dc.contributor.none.fl_str_mv |
Alves, Kele Amaral http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4297620J0 Jacomini, Jose Octavio http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782265U4 Vieira, Luis Alberto Beletti, Marcelo Emílio http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703947D5 Alves, Bênner Geraldo http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4259761D1 |
dc.contributor.author.fl_str_mv |
Antonino, Deize de Cássia |
dc.subject.por.fl_str_mv |
Veterinária Bovino - Reprodução Fertilização in vitro Nanopartículas Folículos pré-antrais Cultivo tridimensional Levitação magnética Nanoparticles Magnetic levitation Preantral follicles Three-dimensional culture CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
topic |
Veterinária Bovino - Reprodução Fertilização in vitro Nanopartículas Folículos pré-antrais Cultivo tridimensional Levitação magnética Nanoparticles Magnetic levitation Preantral follicles Three-dimensional culture CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The aim of this study was to evaluate the development of secondary follicles in the in vitro culture on a three-dimensional system of nanoparticles (gold, iron oxide and poly-L-lysine) associated with magnetic levitation at three concentrations (50, 100 and 200 μL/mL) compared to 2D in vitro culture (control, traditional) for 16 days. Sixty bovine ovaries were collected and the micro dissection of the secondary preantral follicles (200 μm to 400 μm in diameter) was done manually from the fragments of the ovarian cortex. Thereafter, the isolated follicles were individually transferred to micro tubes containing culture medium. For 3D treatments, the nanoparticles were added according to the proposed concentrations. After 12 hours of incubation, the follicles were transferred to 24-well plates with culture medium and cultured for 16 days. At the end of the culture, the oocytes were in vitro matured for 22-24 hours. A total of 213 preantral follicles were selected for in vitro culture and randomly assigned to the treatments. The 200 μL/mL treatment showed a reduction (P < 0.05) in follicular viability only from D12 and at the end of the culture (D16), the highest proportion of viable follicles (P < 0.05) was observed in this group. The treatments with the highest concentrations of nanoparticles had the lowest rates of oocyte extrusion and higher proportion of antrum formation. In addition, within the slow-growing category, a superior percentage of follicles (P < 0.05) was observed in the 200 μL/mL group. The follicles of the control group were 6 times more likely (P < 0.05) of follicular degeneration and/or extrusion when compared to 200 μL/mL group. Additionally, the oocyte viability rate in in vitro maturation was higher (P < 0.05) in the treatment of 200 μL/mL. Furthermore, only the oocytes developed in the 3D culture reached the MI phase after the resumption of meiosis. Finally, it was concluded that the three-dimensional culture system with nanoparticles associated with magnetic levitation promoted the in vitro development of bovine secondary follicles and the concentration of 200 μL/mL obtained the highest proportion of viable follicles with the best rates of antrum formation and maintenance. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-06-13T12:37:11Z 2017-06-13T12:37:11Z 2017-02-24 |
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 |
ANTONINO, Deize de Cássia. Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética. 2017. 71 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Uberlândia, Uberlândia, 2017. DOI http://doi.org/10.14393/ufu.di.2017.222 https://repositorio.ufu.br/handle/123456789/18853 http://doi.org/10.14393/ufu.di.2017.222 |
identifier_str_mv |
ANTONINO, Deize de Cássia. Cultivo in vitro de folículos pré-antrais bovinos associado à nanopartículas e levitação magnética. 2017. 71 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Uberlândia, Uberlândia, 2017. DOI http://doi.org/10.14393/ufu.di.2017.222 |
url |
https://repositorio.ufu.br/handle/123456789/18853 http://doi.org/10.14393/ufu.di.2017.222 |
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 |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Ciências Veterinárias |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Ciências Veterinárias |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
instname_str |
Universidade Federal de Uberlândia (UFU) |
instacron_str |
UFU |
institution |
UFU |
reponame_str |
Repositório Institucional da UFU |
collection |
Repositório Institucional da UFU |
repository.name.fl_str_mv |
Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU) |
repository.mail.fl_str_mv |
diinf@dirbi.ufu.br |
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1805569563007385600 |