Non-Viral Transfection of Human T Lymphocytes

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Riedl, Simon ; Kaiser, Patrick ; Raup, Alexander ; Synatschke, Christopher V. ; Jérôme, Valérie ; Freitag, Ruth:
Non-Viral Transfection of Human T Lymphocytes.
In: Processes. Bd. 6 (2018) Heft 10 . - 188.
ISSN 2227-9717
DOI: https://doi.org/10.3390/pr6100188

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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.