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dc.contributor.authorB. Tinsley, Rogan
dc.contributor.authorJ. Vesey, Melanie
dc.contributor.authorBarati, Shahram
dc.contributor.authorA. Rush, Robert
dc.contributor.authorA. Ferguson, Ian
dc.date.accessioned2017-05-03T16:57:55Z
dc.date.available2017-05-03T16:57:55Z
dc.date.issued2004
dc.date.modified2010-01-07T02:17:05Z
dc.identifier.issn1099498X
dc.identifier.doi10.1002/jgm.584
dc.identifier.urihttp://hdl.handle.net/10072/28086
dc.description.abstractBackground Rational design of gene vectors for therapeutic applications requires understanding of transfection mechanisms. In this study, multiple transfection assays revealed complementary mechanisms between two commonly used transfection agents. This finding was then exploited to produce improved transfection outcomes. Methods and results Rat C6 glial cells, adult rat hippocampal progenitor cells and primary astrocytes were transfected using Lipofectamine (LA) or polyethylenimine (PEI), in vitro. Although LA- and PEI-transfected populations expressed the same total level of transgene product, LA transfected considerably more cells than PEI (approximately 20 vs. 14%). A fluorescently labelled plasmid and time-course analysis, involving both flow cytometry and confocal microscopy, were used to explain this apparent discrepancy. Results showed that LA delivered more plasmid DNA to the cytoplasm and achieved transgene expression in more cells than PEI. In contrast, PEI transfected fewer cells but, on average, produced more transgene product per transfected cell. Conclusions A comparative transfection model was developed to explain these different characteristics. According to this model, transfection is a multistage process with different transfection agents exerting their primary effect at different stages in this process. This model forecast that it should be possible to prepare a chimeric complex with a transfection efficiency that exceeded that achievable with Lipofectamine or polyethylenimine alone. This prediction was tested and shown to hold for glioma cells, primary astrocytes, and adult neural stems cells.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherWiley & Son
dc.publisher.placeUnited States
dc.relation.ispartofpagefrom1023
dc.relation.ispartofpageto1032
dc.relation.ispartofissue9
dc.relation.ispartofjournalThe Journal of Gene Medicine
dc.relation.ispartofvolume6
dc.subject.fieldofresearchRegenerative Medicine (incl. Stem Cells and Tissue Engineering)
dc.subject.fieldofresearchClinical Sciences
dc.subject.fieldofresearchcode100404
dc.subject.fieldofresearchcode1103
dc.titleImproved non-viral transfection of glial and adult neural stem cell lines and of primary astrocytes by combining agents with complementary modes of action
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2004
gro.hasfulltextNo Full Text
gro.griffith.authorFerguson, Ian


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