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Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding
Intracellular delivery of functional macromolecules, such as DNA and RNA, across the cell membrane and into the cytosol, is a critical process in both biology and medicine. Herein, we develop and use microfluidic chips containing post arrays to induce microfluidic vortex shedding, or μVS, for cell m...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397276/ https://www.ncbi.nlm.nih.gov/pubmed/30824814 http://dx.doi.org/10.1038/s41598-019-40147-y |
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author | Jarrell, Justin A. Twite, Amy A. Lau, Katherine H. W. J. Kashani, Moein N. Lievano, Adrian A. Acevedo, Julyana Priest, Craig Nieva, Jorge Gottlieb, David Pawell, Ryan S. |
author_facet | Jarrell, Justin A. Twite, Amy A. Lau, Katherine H. W. J. Kashani, Moein N. Lievano, Adrian A. Acevedo, Julyana Priest, Craig Nieva, Jorge Gottlieb, David Pawell, Ryan S. |
author_sort | Jarrell, Justin A. |
collection | PubMed |
description | Intracellular delivery of functional macromolecules, such as DNA and RNA, across the cell membrane and into the cytosol, is a critical process in both biology and medicine. Herein, we develop and use microfluidic chips containing post arrays to induce microfluidic vortex shedding, or μVS, for cell membrane poration that permits delivery of mRNA into primary human T lymphocytes. We demonstrate transfection with μVS by delivery of a 996-nucleotide mRNA construct encoding enhanced green fluorescent protein (EGFP) and assessed transfection efficiencies by quantifying levels of EGFP protein expression. We achieved high transfection efficiency (63.6 ± 3.44% EGFP + viable cells) with high cell viability (77.3 ± 0.58%) and recovery (88.7 ± 3.21%) in CD3 + T cells 19 hrs after μVS processing. Importantly, we show that processing cells via μVS does not negatively affect cell growth rates or alter cell states. We also demonstrate processing speeds of greater than 2.0 × 10(6) cells s(−1) at volumes ranging from 0.1 to 1.5 milliliters. Altogether, these results highlight the use of μVS as a rapid and gentle delivery method with promising potential to engineer primary human cells for research and clinical applications. |
format | Online Article Text |
id | pubmed-6397276 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-63972762019-03-05 Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding Jarrell, Justin A. Twite, Amy A. Lau, Katherine H. W. J. Kashani, Moein N. Lievano, Adrian A. Acevedo, Julyana Priest, Craig Nieva, Jorge Gottlieb, David Pawell, Ryan S. Sci Rep Article Intracellular delivery of functional macromolecules, such as DNA and RNA, across the cell membrane and into the cytosol, is a critical process in both biology and medicine. Herein, we develop and use microfluidic chips containing post arrays to induce microfluidic vortex shedding, or μVS, for cell membrane poration that permits delivery of mRNA into primary human T lymphocytes. We demonstrate transfection with μVS by delivery of a 996-nucleotide mRNA construct encoding enhanced green fluorescent protein (EGFP) and assessed transfection efficiencies by quantifying levels of EGFP protein expression. We achieved high transfection efficiency (63.6 ± 3.44% EGFP + viable cells) with high cell viability (77.3 ± 0.58%) and recovery (88.7 ± 3.21%) in CD3 + T cells 19 hrs after μVS processing. Importantly, we show that processing cells via μVS does not negatively affect cell growth rates or alter cell states. We also demonstrate processing speeds of greater than 2.0 × 10(6) cells s(−1) at volumes ranging from 0.1 to 1.5 milliliters. Altogether, these results highlight the use of μVS as a rapid and gentle delivery method with promising potential to engineer primary human cells for research and clinical applications. Nature Publishing Group UK 2019-03-01 /pmc/articles/PMC6397276/ /pubmed/30824814 http://dx.doi.org/10.1038/s41598-019-40147-y Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Jarrell, Justin A. Twite, Amy A. Lau, Katherine H. W. J. Kashani, Moein N. Lievano, Adrian A. Acevedo, Julyana Priest, Craig Nieva, Jorge Gottlieb, David Pawell, Ryan S. Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title | Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title_full | Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title_fullStr | Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title_full_unstemmed | Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title_short | Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding |
title_sort | intracellular delivery of mrna to human primary t cells with microfluidic vortex shedding |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6397276/ https://www.ncbi.nlm.nih.gov/pubmed/30824814 http://dx.doi.org/10.1038/s41598-019-40147-y |
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