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Gold Nanoparticle-Mediated Photoporation Enables Delivery of Macromolecules over a Wide Range of Molecular Weights in Human CD4+ T Cells

1
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium
2
HIV Cure Research Center, Department of Internal Medicine, Ghent University, Corneel Heymanslaan 10, 9000 Ghent, Belgium
*
Author to whom correspondence should be addressed.
Crystals 2019, 9(8), 411; https://doi.org/10.3390/cryst9080411
Received: 17 June 2019 / Revised: 31 July 2019 / Accepted: 5 August 2019 / Published: 7 August 2019
(This article belongs to the Special Issue Synthesis and Investigation of Gold Nanoparticles)
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Abstract

The modification of CD4+ T cells with exogenous nucleic acids or proteins is a critical step in several research and therapeutic applications, such as HIV studies and cancer immunotherapies. However, efficient cell transfections are not always easily achieved when working with these primary hard-to-transfect cells. While the modification of T cells is typically performed by viral transduction or electroporation, their use is associated with safety issues or cytotoxicity. Vapor nanobubble (VNB) photoporation with sensitizing gold nanoparticles (AuNPs) has recently emerged as a new technology for safe and flexible cell transfections. In this work, we evaluated the potential of VNB photoporation as a novel technique for the intracellular delivery of macromolecules in primary human CD4+ T cells using fluorescent dextrans as model molecules. Our results show that VNB photoporation enables efficient delivery of fluorescent dextrans of 10 kDa in Jurkat (>60% FD10+ cells) as well as in primary human CD4+ T cells (±40% FD10+ cells), with limited cell toxicity (>70% cell viability). We also demonstrated that the technique allows the delivery of dextrans that are up to 500 kDa in Jurkat cells, suggesting its applicability for the delivery of biological macromolecules with a wide range of molecular weights. Altogether, VNB photoporation represents a promising technique for the universal delivery of macromolecules in view of engineering CD4+ T cells for use in a wide variety of research and therapeutic applications. View Full-Text
Keywords: gold nanoparticles; vapor nanobubble; photoporation; T cells; intracellular delivery; macromolecules gold nanoparticles; vapor nanobubble; photoporation; T cells; intracellular delivery; macromolecules
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Raes, L.; Van Hecke, C.; Michiels, J.; Stremersch, S.; Fraire, J.C.; Brans, T.; Xiong, R.; De Smedt, S.; Vandekerckhove, L.; Raemdonck, K.; Braeckmans, K. Gold Nanoparticle-Mediated Photoporation Enables Delivery of Macromolecules over a Wide Range of Molecular Weights in Human CD4+ T Cells. Crystals 2019, 9, 411.

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