ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS

Detalhes bibliográficos
Autor(a) principal: Alves, Endrigo Gabellini Leonel
Data de Publicação: 2017
Outros Autores: Serakides, Rogéria, Rosado, Isabel Rodrigues, Boeloni, Jankerle Neves, Ocarino, Natalia Melo, Rezende, Cleuza Maria de Faria
Tipo de documento: Artigo
Idioma: por
Título da fonte: Ciência animal brasileira (Online)
Texto Completo: https://revistas.ufg.br/vet/article/view/e-34050
Resumo: The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.Keywords: cell culture; differentiation; regenerative medicine.
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spelling ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGSISOLAMENTO E CULTIVO DE CÉLULAS TRONCO MESENQUIMAIS EXTRAÍDAS DO TECIDO ADIPOSO E DA MEDULA ÓSSEA DE CÃESAnimal Sanitymedicina regenerativacultivo celulardiferenciaçãoThe objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.Keywords: cell culture; differentiation; regenerative medicine.Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães. Palavras-chaves: cultivo celular; diferenciação; medicina regenerativa.Universidade Federal de Goiás2017-07-31info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionPesquisa CientíficaResearchapplication/pdftext/htmlhttps://revistas.ufg.br/vet/article/view/e-3405010.1590/cab18034050Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 18 (2017): Continuous publicationCiência Animal Brasileira / Brazilian Animal Science; v. 18 (2017): Publicação contínua1809-68911518-2797reponame:Ciência animal brasileira (Online)instname:Universidade Federal de Goiás (UFG)instacron:UFGporhttps://revistas.ufg.br/vet/article/view/e-34050/23500https://revistas.ufg.br/vet/article/view/e-34050/23501Alves, Endrigo Gabellini LeonelSerakides, RogériaRosado, Isabel RodriguesBoeloni, Jankerle NevesOcarino, Natalia MeloRezende, Cleuza Maria de Fariainfo:eu-repo/semantics/openAccess2023-06-07T18:48:26Zoai:ojs.revistas.ufg.br:article/34050Revistahttps://revistas.ufg.br/vetPUBhttps://revistas.ufg.br/vet/oai||revistacab@gmail.com1809-68911518-2797opendoar:2024-05-21T19:56:09.339682Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG)true
dc.title.none.fl_str_mv ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
ISOLAMENTO E CULTIVO DE CÉLULAS TRONCO MESENQUIMAIS EXTRAÍDAS DO TECIDO ADIPOSO E DA MEDULA ÓSSEA DE CÃES
title ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
spellingShingle ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
Alves, Endrigo Gabellini Leonel
Animal Sanity
medicina regenerativa
cultivo celular
diferenciação
title_short ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
title_full ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
title_fullStr ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
title_full_unstemmed ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
title_sort ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
author Alves, Endrigo Gabellini Leonel
author_facet Alves, Endrigo Gabellini Leonel
Serakides, Rogéria
Rosado, Isabel Rodrigues
Boeloni, Jankerle Neves
Ocarino, Natalia Melo
Rezende, Cleuza Maria de Faria
author_role author
author2 Serakides, Rogéria
Rosado, Isabel Rodrigues
Boeloni, Jankerle Neves
Ocarino, Natalia Melo
Rezende, Cleuza Maria de Faria
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Alves, Endrigo Gabellini Leonel
Serakides, Rogéria
Rosado, Isabel Rodrigues
Boeloni, Jankerle Neves
Ocarino, Natalia Melo
Rezende, Cleuza Maria de Faria
dc.subject.por.fl_str_mv Animal Sanity
medicina regenerativa
cultivo celular
diferenciação
topic Animal Sanity
medicina regenerativa
cultivo celular
diferenciação
description The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.Keywords: cell culture; differentiation; regenerative medicine.
publishDate 2017
dc.date.none.fl_str_mv 2017-07-31
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Pesquisa Científica
Research
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://revistas.ufg.br/vet/article/view/e-34050
10.1590/cab18034050
url https://revistas.ufg.br/vet/article/view/e-34050
identifier_str_mv 10.1590/cab18034050
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv https://revistas.ufg.br/vet/article/view/e-34050/23500
https://revistas.ufg.br/vet/article/view/e-34050/23501
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
text/html
dc.publisher.none.fl_str_mv Universidade Federal de Goiás
publisher.none.fl_str_mv Universidade Federal de Goiás
dc.source.none.fl_str_mv Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 18 (2017): Continuous publication
Ciência Animal Brasileira / Brazilian Animal Science; v. 18 (2017): Publicação contínua
1809-6891
1518-2797
reponame:Ciência animal brasileira (Online)
instname:Universidade Federal de Goiás (UFG)
instacron:UFG
instname_str Universidade Federal de Goiás (UFG)
instacron_str UFG
institution UFG
reponame_str Ciência animal brasileira (Online)
collection Ciência animal brasileira (Online)
repository.name.fl_str_mv Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG)
repository.mail.fl_str_mv ||revistacab@gmail.com
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