ISOLATION AND CULTURE OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE AND BONE MARROW OF DOGS
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , |
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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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 |
_version_ |
1799874788787748864 |