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Open AccessArticle

Non-Viral Transfection of Human T Lymphocytes

1
Process Biotechnology, University of Bayreuth, 95440 Bayreuth, Germany
2
Agap2—HIQ Consulting GmbH, 60323 Frankfurt am Main, Germany
3
Department Synthesis of Macromolecules, Max Planck Institute for Polymer Research, 55128 Mainz, Germany
*
Author to whom correspondence should be addressed.
Processes 2018, 6(10), 188; https://doi.org/10.3390/pr6100188
Received: 10 September 2018 / Revised: 27 September 2018 / Accepted: 9 October 2018 / Published: 11 October 2018
(This article belongs to the Special Issue Transient Gene Expression for Rapid Protein and Virus-Vector Supply)
The genetic modification of human T lymphocytes with established non-viral methods is inefficient. Linear polyethylenimine (l-PEI), one of the most popular non-viral transfection agents for mammalian cells in general, only achieves transfection rates in the single digit percentage range for these cells. Here, a well-defined 24-armed poly(2-dimethylamino) ethyl methacrylate (PDMAEMA) nanostar (number average of the molecular weight: 755 kDa, polydispersity: <1.21) synthesized via atom transfer radical polymerization (ATRP) from a silsesquioxane initiator core is proposed as alternative. The agent is used to prepare polyplexes with plasmid DNA (pDNA). Under optimal conditions these polyplexes reproducibly transfect >80% of the cells from a human T-cell leukemia cell line (Jurkat cells) at viabilities close to 90%. The agent also promotes pDNA uptake when simply added to a mixture of cells and pDNA. This constitutes a particular promising approach for efficient transient transfection at large scale. Finally, preliminary experiments were carried out with primary T cells from two different donors. Results were again significantly better than for l-PEI, although further research into the response of individual T cells to the transfection agent will be necessary, before either method can be used to routinely transfect primary T lymphocytes. View Full-Text
Keywords: non-viral gene delivery; Jurkat cells; human T lymphocytes; primary cells; poly(2-dimethylamino) ethyl methacrylate; polycation; nanostar non-viral gene delivery; Jurkat cells; human T lymphocytes; primary cells; poly(2-dimethylamino) ethyl methacrylate; polycation; nanostar
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MDPI and ACS Style

Riedl, S.A.B.; Kaiser, P.; Raup, A.; Synatschke, C.V.; Jérôme, V.; Freitag, R. Non-Viral Transfection of Human T Lymphocytes. Processes 2018, 6, 188.

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