Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Detalhes bibliográficos
Autor(a) principal: Carvalho, Armando De M. [UNESP]
Data de Publicação: 2013
Outros Autores: 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]
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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spelling 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
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