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

DOI zum Zitieren der Version auf EPub Bayreuth: https://doi.org/10.15495/EPub_UBT_00006533
URN to cite this document: urn:nbn:de:bvb:703-epub-6533-2

Title data

Keim, Daniel ; Gollner, Katrin ; Gollner, Ulrich ; Jérôme, Valérie ; Freitag, Ruth:
Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors.
In: International Journal of Molecular Sciences. Vol. 22 (2021) Issue 15 . - No. 8239.
ISSN 1422-0067
DOI der Verlagsversion: https://doi.org/10.3390/ijms22158239

Format: PDF
Name: ijms-22-08239-v2.pdf
Version: Published Version
Available under License Creative Commons BY 4.0: Attribution
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Project financing: Deutsche Forschungsgemeinschaft


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.

Further data

Item Type: Article in a journal
Keywords: non-viral gene delivery; human B lymphocytes; primary cells; poly(2-dimethylamino)
ethyl methacrylate; polycation; nano-star
DDC Subjects: 500 Science > 570 Life sciences, biology
600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 600 Technology
600 Technology, medicine, applied sciences > 610 Medicine and health
Institutions of the University: Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Language: English
Originates at UBT: Yes
URN: urn:nbn:de:bvb:703-epub-6533-2
Date Deposited: 22 Jul 2022 04:57
Last Modified: 22 Jul 2022 04:57
URI: https://epub.uni-bayreuth.de/id/eprint/6533


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