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

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: https://doi.org/10.3390/ijms22158239

Official URL: Volltext

Project information

Project title:
Project's official titleProject's id
No information411774929

Project financing: Deutsche Forschungsgemeinschaft

Abstract in another language

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
Refereed: Yes
Keywords: non-viral gene delivery; human B lymphocytes; primary cells; poly(2-dimethylamino)
ethyl methacrylate; polycation; nano-star
Institutions of the University: Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Result of work at the UBT: Yes
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
Date Deposited: 24 Aug 2021 07:21
Last Modified: 24 Aug 2021 07:21
URI: https://eref.uni-bayreuth.de/id/eprint/66832