Abstract
With the inherent antitumor function and unique “off-the-shelf” potential, genetically engineered human natural killer (NK) cells with chimeric antigen receptors (CARs) bear great promise for the treatment of multiple hematological malignancies and solid tumors. Current methods of producing large-scale CAR-NK cells mainly rely on mRNA transfection and viral vector transduction. However, mRNA CAR-NK cells were not stable in CAR expression while viral vector transduction mostly ended up with low efficiency. In this chapter, we described an optimized protocol to generate CAR-NK cells by using the piggyBac transposon system via electroporation and to further expand these engineered CAR-NK cells in a large scale together with artificial antigen-presenting feeder cells. This method can stably engineer human primary NK cells with high efficiency and supply sufficient scale of engineered CAR-NK cells for the future possible clinical applications.
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Acknowledgement
This work was supported by the Singapore Ministry of Health’s National Medical Research Council (NMRC/CIRG/1406/2014; NMRC/OFLCG/003/2018) and the establishment of a scalable capability for autologous clinical cell therapy manufacturing (IAF-PP, H18AHa0001) from the Agency for Science, Technology and Research (A*STAR).
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Du, Z., Zhao, T., Chen, X., Zha, S., Wang, S. (2024). A Nonviral piggyBac Transposon-Mediated Method to Generate Large-Scale CAR-NK Cells from Human Peripheral Blood Primary NK Cells. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_18
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DOI: https://doi.org/10.1007/978-1-0716-3593-3_18
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-3593-3
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