Highly Efficient Transfection of Human Primary T Lymphocytes Using Droplet-Enabled Mechanoporation

Byeongju Joo, Jeongsoo Hur, Gi Beom Kim, Seung Gyu Yun, Aram J. Chung

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)12888-12898
Number of pages11
JournalACS nano
Issue number8
Publication statusPublished - 2021 Aug 24

Bibliographical note

Funding Information:
The authors would like to thank Ms. S. B. Jang and Prof. S.G. Cho at Konkuk University for the preparation of plasmid DNAs, and all members of the Biomicrofluidics Laboratory at Korea University for their technical support and useful discussions. This work was supported by a Korea University grant (K1916951) and the National Research Foundation of Korea (NRF), funded by the Ministry of Education (NRF-2018R1D1A1B07045538) and the Ministry of Science and ICT (No. 2021R1A2C2006224). Provisional patents were filed at the authors’ institution.

Publisher Copyright:
© 2021 American Chemical Society.


  • T cell engineering
  • cell therapy
  • droplet microfluidics
  • droplet squeezing
  • gene delivery
  • intracellular delivery
  • transfection

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)


Dive into the research topics of 'Highly Efficient Transfection of Human Primary T Lymphocytes Using Droplet-Enabled Mechanoporation'. Together they form a unique fingerprint.

Cite this