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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
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Project information
| Project title: |
Project's official title Project's id Open Access Publizieren No information |
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| Project financing: |
Deutsche Forschungsgemeinschaft |
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.
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 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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