Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/169019 |
Resumo: | Objective: The aim of this study has been the isolation and characterization of stem cells from dental pulp (DPSCs) in culture. Methods: The primary DPSCs cultures were obtained from human third molars. Immediately after extraction, the teeth were placed in Dulbecco’s modified Eagle medium (DMEM) culture medium supplemented with fetal bovine serum and antibiotics. In a laminar flow, the pulp was removed from the tooth, incubated with collagenase for 2 hours and placed on a culture plate. Phenotypic characterization and cell pluripotency was performed in the fifth passage (P5). To evaluate the expression of surface markers, the cells were incubated with antibodies against CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105 and HLA-DR antigens. For induction of cell differentiation in vitro, 104 cells/cm2 were plated in 12-well plates and cultivated in appropriate media for osteogenic, adipogenic and condrogencic differentiation after reaching at least 70% confluence. Results: The cells were positive above 95% for characteristic markers of the mesenchymal stem cells CD29, CD44, CD73 and CD90. In contrast, there was a low percentage of positivity (up to 1.1%) for the characteristic markers of hematopoietic cells such as CD14, CD34, CD45, CD184 and HLA-DR. Adipogenic differentiation was visualized by staining lipid vacuoles with Oil Red. Bone differentiation was visualized by staining calcium deposits with Alizarin Red. No chondrogenic differentiation was observed. Conclusions: The isolated cells were adherent to plastic, positive for the characteristic markers of mesenchymal stem cells and, therefore, an alternative source for tissue engineering studies. |
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Luisi, Simone BonatoSant'Ana Filho, ManoelPranke, Patricia Helena Lucas2017-09-28T02:28:32Z20172058-5314http://hdl.handle.net/10183/169019001022190Objective: The aim of this study has been the isolation and characterization of stem cells from dental pulp (DPSCs) in culture. Methods: The primary DPSCs cultures were obtained from human third molars. Immediately after extraction, the teeth were placed in Dulbecco’s modified Eagle medium (DMEM) culture medium supplemented with fetal bovine serum and antibiotics. In a laminar flow, the pulp was removed from the tooth, incubated with collagenase for 2 hours and placed on a culture plate. Phenotypic characterization and cell pluripotency was performed in the fifth passage (P5). To evaluate the expression of surface markers, the cells were incubated with antibodies against CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105 and HLA-DR antigens. For induction of cell differentiation in vitro, 104 cells/cm2 were plated in 12-well plates and cultivated in appropriate media for osteogenic, adipogenic and condrogencic differentiation after reaching at least 70% confluence. Results: The cells were positive above 95% for characteristic markers of the mesenchymal stem cells CD29, CD44, CD73 and CD90. In contrast, there was a low percentage of positivity (up to 1.1%) for the characteristic markers of hematopoietic cells such as CD14, CD34, CD45, CD184 and HLA-DR. Adipogenic differentiation was visualized by staining lipid vacuoles with Oil Red. Bone differentiation was visualized by staining calcium deposits with Alizarin Red. No chondrogenic differentiation was observed. Conclusions: The isolated cells were adherent to plastic, positive for the characteristic markers of mesenchymal stem cells and, therefore, an alternative source for tissue engineering studies.application/pdfengDental, oral and craniofacial research. London. Vol. 3, no. 5 (2017), p. 1-3Polpa dentáriaEngenharia tecidualCélulas-troncoCell cultureHuman dental pulpStem cellsIsolation, immunophenotypic characterization and pluripotency of dental pulp stem cellsEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSORIGINAL001022190.pdf001022190.pdfTexto completo (inglês)application/pdf406382http://www.lume.ufrgs.br/bitstream/10183/169019/1/001022190.pdff913680dc59a9e02fc48f3f1491b0cb9MD51TEXT001022190.pdf.txt001022190.pdf.txtExtracted Texttext/plain15769http://www.lume.ufrgs.br/bitstream/10183/169019/2/001022190.pdf.txtff9f9ea21a8c8e30f05ccff0b0bd277dMD52THUMBNAIL001022190.pdf.jpg001022190.pdf.jpgGenerated Thumbnailimage/jpeg1983http://www.lume.ufrgs.br/bitstream/10183/169019/3/001022190.pdf.jpg3cd57a8baed58f3a8017ca4f36b48375MD5310183/1690192019-06-28 02:35:55.050688oai:www.lume.ufrgs.br:10183/169019Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2019-06-28T05:35:55Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
title |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
spellingShingle |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells Luisi, Simone Bonato Polpa dentária Engenharia tecidual Células-tronco Cell culture Human dental pulp Stem cells |
title_short |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
title_full |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
title_fullStr |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
title_full_unstemmed |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
title_sort |
Isolation, immunophenotypic characterization and pluripotency of dental pulp stem cells |
author |
Luisi, Simone Bonato |
author_facet |
Luisi, Simone Bonato Sant'Ana Filho, Manoel Pranke, Patricia Helena Lucas |
author_role |
author |
author2 |
Sant'Ana Filho, Manoel Pranke, Patricia Helena Lucas |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Luisi, Simone Bonato Sant'Ana Filho, Manoel Pranke, Patricia Helena Lucas |
dc.subject.por.fl_str_mv |
Polpa dentária Engenharia tecidual Células-tronco |
topic |
Polpa dentária Engenharia tecidual Células-tronco Cell culture Human dental pulp Stem cells |
dc.subject.eng.fl_str_mv |
Cell culture Human dental pulp Stem cells |
description |
Objective: The aim of this study has been the isolation and characterization of stem cells from dental pulp (DPSCs) in culture. Methods: The primary DPSCs cultures were obtained from human third molars. Immediately after extraction, the teeth were placed in Dulbecco’s modified Eagle medium (DMEM) culture medium supplemented with fetal bovine serum and antibiotics. In a laminar flow, the pulp was removed from the tooth, incubated with collagenase for 2 hours and placed on a culture plate. Phenotypic characterization and cell pluripotency was performed in the fifth passage (P5). To evaluate the expression of surface markers, the cells were incubated with antibodies against CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105 and HLA-DR antigens. For induction of cell differentiation in vitro, 104 cells/cm2 were plated in 12-well plates and cultivated in appropriate media for osteogenic, adipogenic and condrogencic differentiation after reaching at least 70% confluence. Results: The cells were positive above 95% for characteristic markers of the mesenchymal stem cells CD29, CD44, CD73 and CD90. In contrast, there was a low percentage of positivity (up to 1.1%) for the characteristic markers of hematopoietic cells such as CD14, CD34, CD45, CD184 and HLA-DR. Adipogenic differentiation was visualized by staining lipid vacuoles with Oil Red. Bone differentiation was visualized by staining calcium deposits with Alizarin Red. No chondrogenic differentiation was observed. Conclusions: The isolated cells were adherent to plastic, positive for the characteristic markers of mesenchymal stem cells and, therefore, an alternative source for tissue engineering studies. |
publishDate |
2017 |
dc.date.accessioned.fl_str_mv |
2017-09-28T02:28:32Z |
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2017 |
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001022190 |
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dc.language.iso.fl_str_mv |
eng |
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dc.relation.ispartof.pt_BR.fl_str_mv |
Dental, oral and craniofacial research. London. Vol. 3, no. 5 (2017), p. 1-3 |
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openAccess |
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