Generation of stable cell line by using chitosan as gene delivery system

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dc.authoridŞALVA, EMINE/0000-0002-1159-5850
dc.authoridSalva, Emine/0000-0002-1159-5850
dc.authoridEkentok, Ceyda/0000-0001-7721-8778
dc.authorwosid/AAD-1704-2020
dc.authorwosidŞALVA, EMINE/CAH-3062-2022
dc.authorwosidSalva, Emine/ABI-2766-2020
dc.contributor.authorSalva, Emine
dc.contributor.authorTuran, Suna Ozbas
dc.contributor.authorEkentok, Ceyda
dc.contributor.authorAkbuga, Julide
dc.date.accessioned2024-08-04T20:40:20Z
dc.date.available2024-08-04T20:40:20Z
dc.date.issued2016
dc.departmentİnönü Üniversitesien_US
dc.description.abstractEstablishing stable cell lines are useful tools to study the function of various genes and silence or induce the expression of a gene of interest. Nonviral gene transfer is generally preferred to generate stable cell lines in the manufacturing of recombinant proteins. In this study, we aimed to establish stable recombinant HEK-293 cell lines by transfection of chitosan complexes preparing with pDNA which contain LacZ and GFP genes. Chitosan which is a cationic polymer was used as gene delivery system. Stable HEK-293 cell lines were established by transfection of cells with complexes which were prepared with chitosan and pVitro-2 plasmid vector that contains neomycin drug resistance gene, beta gal and GFP genes. The transfection efficiency was shown with GFP expression in the cells using fluorescence microscopy. Beta gal protein expression in stable cells was examined by beta-galactosidase assay as enzymatically and X-gal staining method as histochemically. Full complexation was shown in the above of 1/1 ratio in the chitosan/pDNA complexes. The highest beta-galactosidase activity was obtained with transfection of chitosan complexes. Beta gal gene expression was 15.17 ng/ml in the stable cells generated by chitosan complexes. In addition, intensive blue color was observed depending on beta gal protein expression in the stable cell line with X-gal staining. We established a stable HEK-293 cell line that can be used for recombinant protein production or gene expression studies by transfecting the gene of interest.en_US
dc.description.sponsorshipMarmara University Scientific Research Project Centre (BAPKO) [SAG-D-140312-0042]en_US
dc.description.sponsorshipThis study was supported Marmara University Scientific Research Project Centre (BAPKO) with grant no: SAG-D-140312-0042en_US
dc.identifier.doi10.1007/s10616-015-9859-8
dc.identifier.endpage1038en_US
dc.identifier.issn0920-9069
dc.identifier.issn1573-0778
dc.identifier.issue4en_US
dc.identifier.pmid26134852en_US
dc.identifier.scopus2-s2.0-84934783726en_US
dc.identifier.scopusqualityQ3en_US
dc.identifier.startpage1033en_US
dc.identifier.urihttps://doi.org/10.1007/s10616-015-9859-8
dc.identifier.urihttps://hdl.handle.net/11616/96857
dc.identifier.volume68en_US
dc.identifier.wosWOS:000380265300042en_US
dc.identifier.wosqualityQ3en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherSpringeren_US
dc.relation.ispartofCytotechnologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectChitosanen_US
dc.subjectBeta galen_US
dc.subjectpDNAen_US
dc.subjectStable cellen_US
dc.titleGeneration of stable cell line by using chitosan as gene delivery systemen_US
dc.typeArticleen_US

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