Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells

Detalhes bibliográficos
Autor(a) principal: De Almeida, Danilo C.
Data de Publicação: 2012
Outros Autores: Santos, Rodrigo Da S., Ribeiro-Paes, João T. [UNESP]
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.
id UNSP_3aeae0b6de79af85c498445b96b8da33
oai_identifier_str oai:repositorio.unesp.br:11449/73638
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling 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
_version_ 1808128812096946176