Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1590/S0100-736X2013000900017 http://hdl.handle.net/11449/109869 |
Resumo: | The objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also cultured in specific media for adipogenic and chondrogenic differentiation. There was no expression of the CD13 marker, but CD44 and CD90 were expressed in all of the passages tested. After 14 days of cell differentiation into adipocytes, lipid droplets were observed upon Oil Red O (ORO) staining. Twenty-one days after chondrogenic differentiation, the cells were stained with Alcian Blue. Although the technique for the isolation of these cells requires improvement, the present study demonstrates the partial characterization of PbMSCs, classifying them as a promising type of progenitor cells for use in equine cell therapy. |
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Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cellsIsolamento e caracterização das células mesenquimais multipotentes estromais derivadas do sangue periférico equinoCaracterização imunofenotípicadiferenciaçãocélulas tronco mesenquimaissangue periféricoequinoImmunophenotypic characterizationdifferentiationmesenchymal stem cellsperipheral bloodequineThe objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also cultured in specific media for adipogenic and chondrogenic differentiation. There was no expression of the CD13 marker, but CD44 and CD90 were expressed in all of the passages tested. After 14 days of cell differentiation into adipocytes, lipid droplets were observed upon Oil Red O (ORO) staining. Twenty-one days after chondrogenic differentiation, the cells were stained with Alcian Blue. Although the technique for the isolation of these cells requires improvement, the present study demonstrates the partial characterization of PbMSCs, classifying them as a promising type of progenitor cells for use in equine cell therapy.O objetivo deste estudo foi isolar, cultivar e caracterizar as células mesenquimais multipotentes estromais derivadas do sangue periférico (SpCTMs) equino. O sangue periférico foi coletado, seguido do isolamento das células mononucleadas utilizando o reagente de gradiente de densidade e o cultivo das células aderentes. Os anticorpos monoclonais mouse anti-horse CD13, mouse anti-horse CD44 e mouse anti-rat CD90 foram utilizados para a caracterização imunofenotípica da superfície das SpCTMs. Estas células também foram cultivadas utilizando meio de cultura específico para a diferenciação adipogênica e condrogênica. Não houve expressão do marcador CD13, mas os marcadores CD44 e CD90 foram expressos em todas as passagens testadas. Após 14 dias da diferenciação das células em adipócitos, gotículas de lipídeos foram observados através da coloração com Oil Red O. Vinte e um dias após a diferenciação condrogênica, as células foram coradas com o Alcian Blue. Embora a técnica de isolamento destas células necessite ser otimizada, o presente estudo demonstra a caracterização parcial das SpCTMs, classificando-as como um tipo de células progenitoras promissoras para o uso na terapia celular em equinos.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Cirurgia e Anestesiologia VeterináriaUnesp Faculdade de Medicina de Botucatu HemocentroUnesp FMVZ Departamento de Clínica VeterináriaUniversidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Cirurgia e Anestesiologia VeterináriaUnesp Faculdade de Medicina de Botucatu HemocentroUnesp FMVZ Departamento de Clínica VeterináriaFAPESP: 09/10670-8Colégio Brasileiro de Patologia Animal - CBPAUniversidade Estadual Paulista (Unesp)Carvalho, Armando De M. [UNESP]Yamada, Ana Lucia M. [UNESP]Martins, Juliana R.b. [UNESP]Maia, Leandro [UNESP]Golim, Marjorie A. [UNESP]Deffune, Elenice [UNESP]Hussni, Carlos Alberto [UNESP]Alves, Ana Liz Garcia [UNESP]2014-10-01T13:08:38Z2014-10-01T13:08:38Z2013-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1151-1154application/pdfhttp://dx.doi.org/10.1590/S0100-736X2013000900017Pesquisa Veterinária Brasileira. Colégio Brasileiro de Patologia Animal - CBPA, v. 33, n. 9, p. 1151-1154, 2013.0100-736Xhttp://hdl.handle.net/11449/10986910.1590/S0100-736X2013000900017S0100-736X2013000900017WOS:000327655200017S0100-736X2013000900017.pdf60209849378498017773733250141398SciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPesquisa Veterinária Brasileira0.385info:eu-repo/semantics/openAccess2024-01-11T06:25:34Zoai:repositorio.unesp.br:11449/109869Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T22:40:09.088444Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells Isolamento e caracterização das células mesenquimais multipotentes estromais derivadas do sangue periférico equino |
title |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
spellingShingle |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells Carvalho, Armando De M. [UNESP] Caracterização imunofenotípica diferenciação células tronco mesenquimais sangue periférico equino Immunophenotypic characterization differentiation mesenchymal stem cells peripheral blood equine |
title_short |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
title_full |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
title_fullStr |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
title_full_unstemmed |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
title_sort |
Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells |
author |
Carvalho, Armando De M. [UNESP] |
author_facet |
Carvalho, Armando De M. [UNESP] Yamada, Ana Lucia M. [UNESP] Martins, Juliana R.b. [UNESP] Maia, Leandro [UNESP] Golim, Marjorie A. [UNESP] Deffune, Elenice [UNESP] Hussni, Carlos Alberto [UNESP] Alves, Ana Liz Garcia [UNESP] |
author_role |
author |
author2 |
Yamada, Ana Lucia M. [UNESP] Martins, Juliana R.b. [UNESP] Maia, Leandro [UNESP] Golim, Marjorie A. [UNESP] Deffune, Elenice [UNESP] Hussni, Carlos Alberto [UNESP] Alves, Ana Liz Garcia [UNESP] |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Carvalho, Armando De M. [UNESP] Yamada, Ana Lucia M. [UNESP] Martins, Juliana R.b. [UNESP] Maia, Leandro [UNESP] Golim, Marjorie A. [UNESP] Deffune, Elenice [UNESP] Hussni, Carlos Alberto [UNESP] Alves, Ana Liz Garcia [UNESP] |
dc.subject.por.fl_str_mv |
Caracterização imunofenotípica diferenciação células tronco mesenquimais sangue periférico equino Immunophenotypic characterization differentiation mesenchymal stem cells peripheral blood equine |
topic |
Caracterização imunofenotípica diferenciação células tronco mesenquimais sangue periférico equino Immunophenotypic characterization differentiation mesenchymal stem cells peripheral blood equine |
description |
The objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also cultured in specific media for adipogenic and chondrogenic differentiation. There was no expression of the CD13 marker, but CD44 and CD90 were expressed in all of the passages tested. After 14 days of cell differentiation into adipocytes, lipid droplets were observed upon Oil Red O (ORO) staining. Twenty-one days after chondrogenic differentiation, the cells were stained with Alcian Blue. Although the technique for the isolation of these cells requires improvement, the present study demonstrates the partial characterization of PbMSCs, classifying them as a promising type of progenitor cells for use in equine cell therapy. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-09-01 2014-10-01T13:08:38Z 2014-10-01T13:08:38Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1590/S0100-736X2013000900017 Pesquisa Veterinária Brasileira. Colégio Brasileiro de Patologia Animal - CBPA, v. 33, n. 9, p. 1151-1154, 2013. 0100-736X http://hdl.handle.net/11449/109869 10.1590/S0100-736X2013000900017 S0100-736X2013000900017 WOS:000327655200017 S0100-736X2013000900017.pdf 6020984937849801 7773733250141398 |
url |
http://dx.doi.org/10.1590/S0100-736X2013000900017 http://hdl.handle.net/11449/109869 |
identifier_str_mv |
Pesquisa Veterinária Brasileira. Colégio Brasileiro de Patologia Animal - CBPA, v. 33, n. 9, p. 1151-1154, 2013. 0100-736X 10.1590/S0100-736X2013000900017 S0100-736X2013000900017 WOS:000327655200017 S0100-736X2013000900017.pdf 6020984937849801 7773733250141398 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Pesquisa Veterinária Brasileira 0.385 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
1151-1154 application/pdf |
dc.publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
dc.source.none.fl_str_mv |
SciELO reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129448688484352 |