vector-free microfluidic platform for intracellular delivery

• Published in: Proceedings of the National Academy of Sciences - PNAS 2013-02, Vol.110 (6), p.2082-2087
• Main Authors: Sharei, Armon, Zoldan, Janet, Adamo, Andrea, Sim, Woo Young, Cho, Nahyun, Jackson, Emily, Mao, Shirley, Schneider, Sabine, Han, Min-Joon, Lytton-Jean, Abigail, Basto, Pamela A., Jhunjhunwala, Siddharth, Lee, Jungmin, Heller, Daniel A., Kang, Jeon Woong, Hartoularos, George C., Kim, Kwang-Soo, Anderson, Daniel G., Langer, Robert, Jensen, Klavs F.
• Format: Article
• Language: English
• Subjects: Animals; B lymphocytes; Biological Sciences; Biomechanical Phenomena; Cell Membrane Permeability; Cell Shape; Cells; Cells, Cultured; Cytosol - metabolism; Dendritic Cells - cytology; Dendritic Cells - metabolism; Dextrans; Diffusion; Drug Delivery Systems; Electric fields; Electroporation; Gene Expression; HeLa Cells; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Materials; Mice; Microfluidic Analytical Techniques; Nanotubes; Nanotubes, Carbon; Peptides; Pluripotent stem cells; Proteins; Proteins - administration & dosage; RNA, Small Interfering - administration & dosage; Stem cells; T lymphocytes; Toxicity; Viability
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1218705110

Intracellular delivery of macromolecules is a challenge in research and therapeutic applications. Existing vector-based and physical methods have limitations, including their reliance on exogenous materials or electrical fields, which can lead to toxicity or off-target effects. We describe a microfluidic approach to delivery in which cells are mechanically deformed as they pass through a constriction 30–80% smaller than the cell diameter. The resulting controlled application of compression and shear forces results in the formation of transient holes that enable the diffusion of material from the surrounding buffer into the cytosol. The method has demonstrated the ability to deliver a range of material, such as carbon nanotubes, proteins, and siRNA, to 11 cell types, including embryonic stem cells and immune cells. When used for the delivery of transcription factors, the microfluidic devices produced a 10-fold improvement in colony formation relative to electroporation and cell-penetrating peptides. Indeed, its ability to deliver structurally diverse materials and its applicability to difficult-to-transfect primary cells indicate that this method could potentially enable many research and clinical applications.