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Processes 2018, 6(10), 188; https://doi.org/10.3390/pr6100188

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.
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)
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Abstract

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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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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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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