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Published on: January 21, 2021
Nanoinjection: A Platform for Innovation in Ex Vivo Cell Engineering
Yaping Chen1,2, Ali-Reza Shokouhi2, Nicolas H Voelcker2,3,4
1Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), Institute of Aging, Key Laboratory of Alzheimer's Disease of Zhejiang Province, Zhejiang Provincial Clinical Research Center for Mental Disorders, The Affiliated Wenzhou Kangning Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325035, P.R. China.
Nanoinjection offers a precise method for delivering therapeutic cargos into human cells, including for engineering chimeric antigen receptor (CAR)-T cells. This advanced technique minimizes cell damage and manufacturing complexities for improved ex vivo cell therapies.
Area of Science:
- Cellular and Molecular Engineering
- Biotechnology and Bioengineering
- Immunotherapy
Background:
- Intracellular delivery of therapeutic cargos like gene-editing tools and nucleic acids is crucial but challenging for cell function.
- Chimeric antigen receptor (CAR)-T cell therapy has shown success in treating blood cancers, but faces manufacturing and safety hurdles.
- Current methods like electroporation (EP) can cause cell damage and limit cargo viability.
Purpose of the Study:
- To introduce nanoinjection as an advanced physical delivery route for intracellular cargo transport.
- To highlight nanoinjection's potential in ex vivo cell engineering, particularly for CAR-T cell generation.
- To compare mechanical and electroactive nanoinjection methods and their advantages over conventional electroporation.
Main Methods:
- Nanoinjection utilizes nanoneedles (NNs) for direct intracellular delivery of diverse therapeutic cargos.
- Mechanical nanoinjection applies direct force for membrane perturbation and cargo transport.
- Electroactive nanoinjection couples NNs with an electric field for nanoscale electroporation, reducing voltage and cell damage.
Main Results:
- Nanoinjection efficiently delivers cargos into various cell types, including primary human T cells, with minimal perturbation and toxicity.
- The technique offers high throughput and resolution, preserving cargo integrity for intracellular function.
- Electroactive nanoinjection significantly reduces the applied voltage compared to bulk EP, minimizing post-procedure cell and cargo damage.
Conclusions:
- Nanoinjection presents a promising platform for ex vivo cell engineering, overcoming limitations of traditional methods.
- This approach facilitates the generation of advanced cell therapies, such as CAR-T cells, with enhanced efficacy and safety.
- Nanoinjection's adaptability supports applications in immunomodulation, mechanotransduction, and cell state sampling.
