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Article

Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors

1
Process Biotechnology, University of Bayreuth, 95447 Bayreuth, Germany
2
Praxis am Schießgraben, Schießgraben 21, 95326 Kulmbach, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Thierry Bordet
Int. J. Mol. Sci. 2021, 22(15), 8239; https://doi.org/10.3390/ijms22158239
Received: 14 July 2021 / Revised: 29 July 2021 / Accepted: 29 July 2021 / Published: 30 July 2021
(This article belongs to the Special Issue Non-viral Gene Delivery: Hurdles and Promises)
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. View Full-Text
Keywords: non-viral gene delivery; human B lymphocytes; primary cells; poly(2-dimethylamino) ethyl methacrylate; polycation; nano-star non-viral gene delivery; human B lymphocytes; primary cells; poly(2-dimethylamino) ethyl methacrylate; polycation; nano-star
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MDPI and ACS Style

Keim, D.; Gollner, K.; Gollner, U.; Jérôme, V.; Freitag, R. Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors. Int. J. Mol. Sci. 2021, 22, 8239. https://doi.org/10.3390/ijms22158239

AMA Style

Keim D, Gollner K, Gollner U, Jérôme V, Freitag R. Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors. International Journal of Molecular Sciences. 2021; 22(15):8239. https://doi.org/10.3390/ijms22158239

Chicago/Turabian Style

Keim, Daniel, Katrin Gollner, Ulrich Gollner, Valérie Jérôme, and Ruth Freitag. 2021. "Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors" International Journal of Molecular Sciences 22, no. 15: 8239. https://doi.org/10.3390/ijms22158239

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