Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/jspui/handle/123456789/17840 |
Resumo: | Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies |
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Antunes, Fernanda Ginanihttp://lattes.cnpq.br/4031509110803993http://lattes.cnpq.br/5004397230198722Santos, Jean Nunes doshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4728907T1Souza, Lélia Batista dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787927Z7&dataRevisao=nullBarbosa, Carlos Augusto Galvão2014-12-17T15:43:54Z2014-08-202014-12-17T15:43:54Z2013-02-14ANTUNES, Fernanda Ginani. Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação. 2013. 61 f. Dissertação (Mestrado em Saúde Pública) - Universidade Federal do Rio Grande do Norte, Natal, 2013.https://repositorio.ufrn.br/jspui/handle/123456789/17840Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studiesCélulas-tronco da polpa dental humana têm sido amplamente investigadas em razão da sua capacidade de diferenciar-se tanto em células dentais quanto não dentais, com potencial de utilização em terapias envolvendo a engenharia de tecidos. A técnica de criopreservação celular representa uma alternativa viável para a conservação dessas células, já que cessa reversivelmente, de forma controlada, todas as suas funções biológicas em uma temperatura ultra-baixa. O presente estudo teve como objetivo avaliar, através de experimentos in vitro, a influência de um protocolo de criopreservação na atividade biológica de células-tronco da polpa de dentes humanos d ecíduos esfoliados (SHED). Células obtidas da polpa de três dentes decíduos em estágio final de esfoliação ou com exodontia indicada foram expandidas em meio de cultivo α-MEM suplementado com antibióticos e 15% de soro fetal bovino. No segundo subcultivo (P2), um grupo de células foi submetido a criopreservação por 30 dias em DMSO diluído a 10% em soro fetal bovino, a 80ºC negativos, enquanto o restante seguiu em condições normais de cultivo. A proliferação celular em ambos os grupos (criopreservado e não criopreservado) foi avaliada através do método de coloração por azul de Tripan nos intervalos de 24, 48 e 72 horas após o plaqueamento. Nestes mesmos intervalos foi realizada a análise do ciclo celular das SHEDs submetidas ou não ao protocolo de criopreservação. Os eventos relacionados à morte celular foram analisados através da expressão de Anexina V e PI em citometria de fluxo, nos intervalos de 24 e 72 horas. A presença de alterações morfológicas nucleares foi avaliada através da marcação por DAPI no intervalo de 72 horas. Observou-se que ambos os grupos exibiram uma curva de proliferação celular ascendente, sem alterações consideráveis na viabilidade celular ao longo do experimento. A distribuição das células nas fases do ciclo celular foi coerente com células em proliferação nos dois grupos. Não foram observados danos morfológicos nucleares no intervalo final do experimento . Deste modo, conclui-se que o protocolo de criopreservação proposto é eficiente para o armazenamento do tipo celular estudado, permitindo a sua utilização em futuros estudos experimentaisCoordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em Saúde ColetivaUFRNBRSaúde PúblicaPolpa dental. Células-tronco. Criopreservação. Cultivo celularDental pulp. Stem cells. Cryopreservation. Cell cultureCNPQ::CIENCIAS DA SAUDE::SAUDE COLETIVAAtividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservaçãoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRNinstname:Universidade Federal do Rio Grande do Norte (UFRN)instacron:UFRNORIGINALFernandaGA_DISSERT.pdfapplication/pdf1532248https://repositorio.ufrn.br/bitstream/123456789/17840/1/FernandaGA_DISSERT.pdf8b25b3bf5e0afc796607cf138b0a9885MD51TEXTFernandaGA_DISSERT.pdf.txtFernandaGA_DISSERT.pdf.txtExtracted texttext/plain90925https://repositorio.ufrn.br/bitstream/123456789/17840/6/FernandaGA_DISSERT.pdf.txt159e4bca1fd7d7679b5db1aef3484d60MD56THUMBNAILFernandaGA_DISSERT.pdf.jpgFernandaGA_DISSERT.pdf.jpgIM Thumbnailimage/jpeg2293https://repositorio.ufrn.br/bitstream/123456789/17840/7/FernandaGA_DISSERT.pdf.jpg42786b6b02321e5885429eddd0e27102MD57123456789/178402017-11-04 16:16:53.353oai:https://repositorio.ufrn.br:123456789/17840Repositório de PublicaçõesPUBhttp://repositorio.ufrn.br/oai/opendoar:2017-11-04T19:16:53Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)false |
dc.title.por.fl_str_mv |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
title |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
spellingShingle |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação Antunes, Fernanda Ginani Polpa dental. Células-tronco. Criopreservação. Cultivo celular Dental pulp. Stem cells. Cryopreservation. Cell culture CNPQ::CIENCIAS DA SAUDE::SAUDE COLETIVA |
title_short |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
title_full |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
title_fullStr |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
title_full_unstemmed |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
title_sort |
Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação |
author |
Antunes, Fernanda Ginani |
author_facet |
Antunes, Fernanda Ginani |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/4031509110803993 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/5004397230198722 |
dc.contributor.referees1.pt_BR.fl_str_mv |
Santos, Jean Nunes dos |
dc.contributor.referees1ID.por.fl_str_mv |
|
dc.contributor.referees1Lattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4728907T1 |
dc.contributor.referees2.pt_BR.fl_str_mv |
Souza, Lélia Batista de |
dc.contributor.referees2Lattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787927Z7&dataRevisao=null |
dc.contributor.author.fl_str_mv |
Antunes, Fernanda Ginani |
dc.contributor.advisor1.fl_str_mv |
Barbosa, Carlos Augusto Galvão |
contributor_str_mv |
Barbosa, Carlos Augusto Galvão |
dc.subject.por.fl_str_mv |
Polpa dental. Células-tronco. Criopreservação. Cultivo celular |
topic |
Polpa dental. Células-tronco. Criopreservação. Cultivo celular Dental pulp. Stem cells. Cryopreservation. Cell culture CNPQ::CIENCIAS DA SAUDE::SAUDE COLETIVA |
dc.subject.eng.fl_str_mv |
Dental pulp. Stem cells. Cryopreservation. Cell culture |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS DA SAUDE::SAUDE COLETIVA |
description |
Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies |
publishDate |
2013 |
dc.date.issued.fl_str_mv |
2013-02-14 |
dc.date.accessioned.fl_str_mv |
2014-12-17T15:43:54Z |
dc.date.available.fl_str_mv |
2014-08-20 2014-12-17T15:43:54Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
ANTUNES, Fernanda Ginani. Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação. 2013. 61 f. Dissertação (Mestrado em Saúde Pública) - Universidade Federal do Rio Grande do Norte, Natal, 2013. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/jspui/handle/123456789/17840 |
identifier_str_mv |
ANTUNES, Fernanda Ginani. Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação. 2013. 61 f. Dissertação (Mestrado em Saúde Pública) - Universidade Federal do Rio Grande do Norte, Natal, 2013. |
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https://repositorio.ufrn.br/jspui/handle/123456789/17840 |
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por |
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Universidade Federal do Rio Grande do Norte |
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UFRN |
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Universidade Federal do Rio Grande do Norte |
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