Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.11606/issn.2176-7262.v45i4p428-435 http://hdl.handle.net/11449/73638 |
Resumo: | Model of study: Experimental study. Introduction: Recently, stem cell research has generated great interest due to its applicability in regenerative medicine. Bone marrow is considered the most important source of adult stem cells and the establishment of new methods towards gene expression analysis regarding stem cells has become necessary. Thus Differential Display Reverse Transcription Polymerase Chain Reaction (DDRT-PCR) may be an accessible tool to investigate small differences in the gene expression of different stem cells in distinct situations. Aim: In the present study, we investigated the exequibility of DDRT-PCR to identify differences in global gene expression of mice bone marrow cells under two conditions. Methods: First, bone marrow cells were isolated fresh and a part was cultivated during one week without medium replacement. Afterwards, both bone marrow cells (fresh and cultivated) were submitted to gene expression analyses by DDRT-PCR. Results: Initially, it was possible to observe in one week-cultured bone marrow cells, changes in morphology (oval cells to fibroblastic-like cells) and protein profile, which was seen through differences in band distribution in SDS-Page gels. Finally through gene expression analysis, we detected three bands (1300, 1000 and 225 bp) exclusively expressed in the fresh bone marrow group and two bands (400 and 300 bp) expressed specifically in the cultivated bone marrow cell group. Conclusions: In summary, the DDRT-PCR method was proved efficient towards the identification of small differences in gene expression of bone marrow cells in two defined conditions. Thus, we expect that DDRT-PCR can be fast and efficiently designed to analyze differential gene expression in several stem cell types under distinct conditions. |
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Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cellsBone Marrow CellsDDRT-PCRGene Expressionanimal cellbone marrow cellcell culturecell isolationcell structuredifferential display reverse transcription polymerase chain reactionfibroblastgene expressiongene expression profilingmousenonhumanpolyacrylamide gel electrophoresisreverse transcription polymerase chain reactionModel of study: Experimental study. Introduction: Recently, stem cell research has generated great interest due to its applicability in regenerative medicine. Bone marrow is considered the most important source of adult stem cells and the establishment of new methods towards gene expression analysis regarding stem cells has become necessary. Thus Differential Display Reverse Transcription Polymerase Chain Reaction (DDRT-PCR) may be an accessible tool to investigate small differences in the gene expression of different stem cells in distinct situations. Aim: In the present study, we investigated the exequibility of DDRT-PCR to identify differences in global gene expression of mice bone marrow cells under two conditions. Methods: First, bone marrow cells were isolated fresh and a part was cultivated during one week without medium replacement. Afterwards, both bone marrow cells (fresh and cultivated) were submitted to gene expression analyses by DDRT-PCR. Results: Initially, it was possible to observe in one week-cultured bone marrow cells, changes in morphology (oval cells to fibroblastic-like cells) and protein profile, which was seen through differences in band distribution in SDS-Page gels. Finally through gene expression analysis, we detected three bands (1300, 1000 and 225 bp) exclusively expressed in the fresh bone marrow group and two bands (400 and 300 bp) expressed specifically in the cultivated bone marrow cell group. Conclusions: In summary, the DDRT-PCR method was proved efficient towards the identification of small differences in gene expression of bone marrow cells in two defined conditions. Thus, we expect that DDRT-PCR can be fast and efficiently designed to analyze differential gene expression in several stem cell types under distinct conditions.Mestre em Ciěncias Médicas Universidade Federal de São Paulo Departamento de Medicina (UNIFESP/EPM), São PauloMestre em Biologia Celular e Molecular Universidade de São Paulo Departamento de Genética (USP/FMRP), Ribeirão Preto-SPUniversidade Estadual Paulista Júlio de Mesquita Filho Departamento de Ciěncias Biológicas (UNESP), Assis-SPLICE Laboratory/UNIFESP,Vila Clementino, Rua Pedro de Toledo, 669, São Paulo 04039-003Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Ciěncias Biológicas (UNESP), Assis-SPUniversidade Federal de São Paulo (UNIFESP)Universidade de São Paulo (USP)Universidade Estadual Paulista (Unesp)De Almeida, Danilo C.Santos, Rodrigo Da S.Ribeiro-Paes, João T. [UNESP]2014-05-27T11:27:05Z2014-05-27T11:27:05Z2012-10-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article424-431application/pdfhttp://dx.doi.org/10.11606/issn.2176-7262.v45i4p428-435Medicina (Brazil), v. 45, n. 4, p. 424-431, 2012.0076-60462176-7262http://hdl.handle.net/11449/7363810.11606/issn.2176-7262.v45i4p428-4352-s2.0-848754096552-s2.0-84875409655.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMedicina (Brazil)0,1250,125info:eu-repo/semantics/openAccess2024-06-13T17:38:20Zoai:repositorio.unesp.br:11449/73638Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:26:30.574100Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
title |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
spellingShingle |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells De Almeida, Danilo C. Bone Marrow Cells DDRT-PCR Gene Expression animal cell bone marrow cell cell culture cell isolation cell structure differential display reverse transcription polymerase chain reaction fibroblast gene expression gene expression profiling mouse nonhuman polyacrylamide gel electrophoresis reverse transcription polymerase chain reaction |
title_short |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
title_full |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
title_fullStr |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
title_full_unstemmed |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
title_sort |
Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells |
author |
De Almeida, Danilo C. |
author_facet |
De Almeida, Danilo C. Santos, Rodrigo Da S. Ribeiro-Paes, João T. [UNESP] |
author_role |
author |
author2 |
Santos, Rodrigo Da S. Ribeiro-Paes, João T. [UNESP] |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Universidade de São Paulo (USP) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
De Almeida, Danilo C. Santos, Rodrigo Da S. Ribeiro-Paes, João T. [UNESP] |
dc.subject.por.fl_str_mv |
Bone Marrow Cells DDRT-PCR Gene Expression animal cell bone marrow cell cell culture cell isolation cell structure differential display reverse transcription polymerase chain reaction fibroblast gene expression gene expression profiling mouse nonhuman polyacrylamide gel electrophoresis reverse transcription polymerase chain reaction |
topic |
Bone Marrow Cells DDRT-PCR Gene Expression animal cell bone marrow cell cell culture cell isolation cell structure differential display reverse transcription polymerase chain reaction fibroblast gene expression gene expression profiling mouse nonhuman polyacrylamide gel electrophoresis reverse transcription polymerase chain reaction |
description |
Model of study: Experimental study. Introduction: Recently, stem cell research has generated great interest due to its applicability in regenerative medicine. Bone marrow is considered the most important source of adult stem cells and the establishment of new methods towards gene expression analysis regarding stem cells has become necessary. Thus Differential Display Reverse Transcription Polymerase Chain Reaction (DDRT-PCR) may be an accessible tool to investigate small differences in the gene expression of different stem cells in distinct situations. Aim: In the present study, we investigated the exequibility of DDRT-PCR to identify differences in global gene expression of mice bone marrow cells under two conditions. Methods: First, bone marrow cells were isolated fresh and a part was cultivated during one week without medium replacement. Afterwards, both bone marrow cells (fresh and cultivated) were submitted to gene expression analyses by DDRT-PCR. Results: Initially, it was possible to observe in one week-cultured bone marrow cells, changes in morphology (oval cells to fibroblastic-like cells) and protein profile, which was seen through differences in band distribution in SDS-Page gels. Finally through gene expression analysis, we detected three bands (1300, 1000 and 225 bp) exclusively expressed in the fresh bone marrow group and two bands (400 and 300 bp) expressed specifically in the cultivated bone marrow cell group. Conclusions: In summary, the DDRT-PCR method was proved efficient towards the identification of small differences in gene expression of bone marrow cells in two defined conditions. Thus, we expect that DDRT-PCR can be fast and efficiently designed to analyze differential gene expression in several stem cell types under distinct conditions. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-10-01 2014-05-27T11:27:05Z 2014-05-27T11:27:05Z |
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/10.11606/issn.2176-7262.v45i4p428-435 Medicina (Brazil), v. 45, n. 4, p. 424-431, 2012. 0076-6046 2176-7262 http://hdl.handle.net/11449/73638 10.11606/issn.2176-7262.v45i4p428-435 2-s2.0-84875409655 2-s2.0-84875409655.pdf |
url |
http://dx.doi.org/10.11606/issn.2176-7262.v45i4p428-435 http://hdl.handle.net/11449/73638 |
identifier_str_mv |
Medicina (Brazil), v. 45, n. 4, p. 424-431, 2012. 0076-6046 2176-7262 10.11606/issn.2176-7262.v45i4p428-435 2-s2.0-84875409655 2-s2.0-84875409655.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Medicina (Brazil) 0,125 0,125 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
424-431 application/pdf |
dc.source.none.fl_str_mv |
Scopus 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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1808128812096946176 |