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

dc.contributor.authorDe Almeida, Danilo C.
dc.contributor.authorSantos, Rodrigo Da S.
dc.contributor.authorRibeiro-Paes, João T. [UNESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-27T11:27:05Z
dc.date.available2014-05-27T11:27:05Z
dc.date.issued2012-10-01
dc.description.abstractModel 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.en
dc.description.affiliationMestre em Ciěncias Médicas Universidade Federal de São Paulo Departamento de Medicina (UNIFESP/EPM), São Paulo
dc.description.affiliationMestre em Biologia Celular e Molecular Universidade de São Paulo Departamento de Genética (USP/FMRP), Ribeirão Preto-SP
dc.description.affiliationUniversidade Estadual Paulista Júlio de Mesquita Filho Departamento de Ciěncias Biológicas (UNESP), Assis-SP
dc.description.affiliationLICE Laboratory/UNIFESP,Vila Clementino, Rua Pedro de Toledo, 669, São Paulo 04039-003
dc.description.affiliationUnespUniversidade Estadual Paulista Júlio de Mesquita Filho Departamento de Ciěncias Biológicas (UNESP), Assis-SP
dc.format.extent424-431
dc.identifierhttp://dx.doi.org/10.11606/issn.2176-7262.v45i4p428-435
dc.identifier.citationMedicina (Brazil), v. 45, n. 4, p. 424-431, 2012.
dc.identifier.doi10.11606/issn.2176-7262.v45i4p428-435
dc.identifier.file2-s2.0-84875409655.pdf
dc.identifier.issn0076-6046
dc.identifier.issn2176-7262
dc.identifier.scopus2-s2.0-84875409655
dc.identifier.urihttp://hdl.handle.net/11449/73638
dc.language.isoeng
dc.relation.ispartofMedicina (Brazil)
dc.relation.ispartofsjr0,125
dc.relation.ispartofsjr0,125
dc.rights.accessRightsAcesso abertopt
dc.sourceScopus
dc.subjectBone Marrow Cells
dc.subjectDDRT-PCR
dc.subjectGene Expression
dc.subjectanimal cell
dc.subjectbone marrow cell
dc.subjectcell culture
dc.subjectcell isolation
dc.subjectcell structure
dc.subjectdifferential display reverse transcription polymerase chain reaction
dc.subjectfibroblast
dc.subjectgene expression
dc.subjectgene expression profiling
dc.subjectmouse
dc.subjectnonhuman
dc.subjectpolyacrylamide gel electrophoresis
dc.subjectreverse transcription polymerase chain reaction
dc.titleExequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cellsen
dc.typeArtigopt
dcterms.licensehttp://www.revistas.usp.br/rmrp/about/editorialPolicies#openAccessPolicy
dspace.entity.typePublication
relation.isDepartmentOfPublication4a016e93-a452-4c24-b800-ecc2ea22a1fd
relation.isDepartmentOfPublication.latestForDiscovery4a016e93-a452-4c24-b800-ecc2ea22a1fd
relation.isOrgUnitOfPublicationc3f68528-5ea8-4b32-a9f4-3cfbd4bba64d
relation.isOrgUnitOfPublication.latestForDiscoveryc3f68528-5ea8-4b32-a9f4-3cfbd4bba64d
unesp.campusUniversidade Estadual Paulista (UNESP), Faculdade de Ciências e Letras, Assispt
unesp.departmentCiências Biológicas - FCLASpt

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