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Cited 26 time in webofscience Cited 31 time in scopus
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Highly Efficient Transfection of Human Primary T Lymphocytes Using Droplet-Enabled Mechanoporation

Authors
Joo, ByeongjuHur, JeongsooKim, Gi-BeomYun, Seung GyuChung, Aram J.
Issue Date
24-Aug-2021
Publisher
American Chemical Society
Keywords
intracellular delivery; gene delivery; T cell engineering; droplet microfluidics; droplet squeezing; transfection; cell therapy
Citation
ACS Nano, v.15, no.8, pp 12888 - 12898
Pages
11
Indexed
SCIE
SCOPUS
Journal Title
ACS Nano
Volume
15
Number
8
Start Page
12888
End Page
12898
URI
https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/54347
DOI
10.1021/acsnano.0c10473
ISSN
1936-0851
1936-086X
Abstract
Whole-cell-based therapy has been extensively used as an effective disease treatment approach, and it has rapidly changed the therapeutic paradigm. To fully accommodate this shift, advances in genome modification and cell reprogramming methodologies are critical. Traditionally, molecular tools such as viral and polymer nanocarriers and electroporation have been the norm for internalizing external biomolecules into cells for cellular engineering. However, these approaches are not fully satisfactory considering their cytotoxicity, high cost, low scalability, and/or inconsistent and ineffective delivery and transfection. To address these challenges, we present an approach that leverages droplet microfluidics with cell mechanoporation, bringing intracellular delivery to the next level. In our approach, cells and external cargos such as mRNAs and plasmid DNAs are coencapsulated into droplets, and as they pass through a series of narrow constrictions, the cell membrane is mechanically permeabilized where the cargos in the vicinity are internalized via convective solution exchange enhanced by recirculation flows developed in the droplets. Using this principle, we demonstrated a high level of functional macromolecule delivery into various immune cells, including human primary T cells. By utilizing droplets, the cargo consumption was drastically reduced, and near-zero clogging was realized. Furthermore, high scalability without sacrificing cell viability and superior delivery over state-of-the-art methods and benchtop techniques were demonstrated. Notably, the droplet-based intracellular delivery strategy presented here can be further applied to other mechanoporation microfluidic techniques, highlighting its potential for cellular engineering and cell-based therapies.
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Yun, Seung Gyu
Anam Hospital (Department of Laboratory Medicine, Anam Hospital)
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