TY  - JOUR
A1  - Moradian, Hanieh
A1  - Roch, Toralf
A1  - Lendlein, Andreas
A1  - Gossen, Manfred
T1  - mRNA transfection-induced activation of primary human monocytes and macrophages
T2  - Scientific reports
N2  - Monocytes and macrophages are key players in maintaining immune homeostasis. Identifying strategies to manipulate their functions via gene delivery is thus of great interest for immunological research and biomedical applications. We set out to establish conditions for mRNA transfection in hard-to-transfect primary human monocytes and monocyte-derived macrophages due to the great potential of gene expression from in vitro transcribed mRNA for modulating cell phenotypes. mRNA doses, nucleotide modifications, and different carriers were systematically explored in order to optimize high mRNA transfer rates while minimizing cell stress and immune activation. We selected three commercially available mRNA transfection reagents including liposome and polymer-based formulations, covering different application spectra. Our results demonstrate that liposomal reagents can particularly combine high gene transfer rates with only moderate immune cell activation. For the latter, use of specific nucleotide modifications proved essential. In addition to improving efficacy of gene transfer, our findings address discrete aspects of innate immune activation using cytokine and surface marker expression, as well as cell viability as key readouts to judge overall transfection efficiency. The impact of this study goes beyond optimizing transfection conditions for immune cells, by providing a framework for assessing new gene carrier systems for monocyte and macrophage, tailored to specific applications.
KW  - sirna transfection
KW  - mediated delivery
KW  - gene delivery
KW  - efficient
KW  - immunogenicity
KW  - lipoplexes
KW  - cells
KW  - therapeutics
KW  - polarization
KW  - pathways
Y1  - 2020
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/61351
SN  - 2045-2322
VL  - 10
IS  - 1
SP  - 1
EP  - 15
PB  - Springer Nature
CY  - London
ER  -