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Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform
Published on: November 7, 2013
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High-Throughput and Efficient Intracellular Delivery Method via a Vibration-Assisted Nanoneedle/Microfluidic
1Department of Mechanical Engineering, Tsinghua University, Beijing 100084, P.R. China.
ACS Nano
|December 8, 2022
Summary
A novel nanoneedle/microfluidic system achieves high-throughput intracellular delivery for tumor immunotherapy. This method efficiently modifies difficult-to-transfect immune cells, like natural killer cells, with minimal damage for enhanced therapeutic potential.
Area of Science:
- Biotechnology
- Cellular Engineering
- Immunotherapy
Background:
- Existing intracellular delivery methods struggle with efficiency, throughput, and cell damage, particularly for suspension immune cells like natural killer cells.
- Natural killer cells are crucial for tumor immunotherapy but are notoriously resistant to genetic modification and transfection.
Purpose of the Study:
- To develop a high-throughput, low-damage system for efficient intracellular delivery and genetic modification of suspension immune cells.
- To overcome the limitations of current transfection techniques for applications in cellular immunotherapy.
Main Methods:
- A vibration-assisted nanoneedle/microfluidic composite system was designed for rapid, large-area cell perforation and detachment.
- Utilized self-assembled nanoneedle arrays within microchannels to facilitate continuous, high-throughput delivery of molecules like Cas9 ribonucleoprotein complexes (Cas9/RNPs).
- Demonstrated the system's capability for gene editing, specifically creating CD96 knockout NK-92 cells.
Main Results:
- Achieved up to 98% delivery efficiency in difficult-to-transfect immune cells with approximately 80% cell viability.
- Demonstrated a throughput at the mL/min level, significantly exceeding conventional methods.
- Successfully generated gene-edited NK-92 cells with enhanced anti-tumor immunity by reversing exhaustion.
Conclusions:
- The developed nanoneedle/microfluidic system offers a highly efficient, high-throughput, and low-damage solution for intracellular delivery.
- This technology holds significant potential for advancing cellular immunotherapy, particularly for engineering natural killer cells for clinical applications.
