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• W3189611252 endingPage "8239" @default.
• W3189611252 startingPage "8239" @default.
• W3189611252 abstract "Although the development of gene delivery systems based on non-viral vectors is advancing, it remains a challenge to deliver plasmid DNA into human blood cells. The current “gold standard”, namely linear polyethyleneimine (l-PEI 25 kDa), in particular, is unable to produce transgene expression levels >5% in primary human B lymphocytes. Here, it is demonstrated that a well-defined 24-armed poly(2-dimethylamino) ethyl methacrylate (PDMAEMA, 755 kDa) nano-star is able to reproducibly elicit high transgene expression (40%) at sufficient residual viability (69%) in primary human B cells derived from tonsillar tissue. Moreover, our results indicate that the length of the mitogenic stimulation prior to transfection is an important parameter that must be established during the development of the transfection protocol. In our hands, four days of stimulation with rhCD40L post-thawing led to the best transfection results in terms of TE and cell survival. Most importantly, our data argue for an impact of the B cell subsets on the transfection outcomes, underlining that the complexity and heterogeneity of a given B cell population pre- and post-transfection is a critical parameter to consider in the multiparametric approach required for the implementation of the transfection protocol." @default.
• W3189611252 created "2021-08-16" @default.
• W3189611252 creator A5030045839 @default.
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• W3189611252 date "2021-07-30" @default.
• W3189611252 modified "2023-10-17" @default.
• W3189611252 title "Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors" @default.
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• W3189611252 doi "https://doi.org/10.3390/ijms22158239" @default.
• W3189611252 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8347318" @default.
• W3189611252 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34361005" @default.
• W3189611252 hasPublicationYear "2021" @default.
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