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bioRxiv · 10.1101/2021.12.28.474351

Assessment of chimeric antigen receptor T (CAR-T) cytotoxicity by droplet microfluidics in vitro

Wong, K. U.·Shi, J.·Li, P.·Wang, H.·Jia, Y.·Deng, C.·Jiang, L.·Wong, A. H.-H.
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Abstract

Chimeric antigen receptor T (CAR-T) cells are cytotoxic T cells engineered to specifically kill cancer cells expressing specific target receptor(s). Prior CAR-T efficacy tests include CAR expression analysis by qPCR or ELISA, in vitro measurement of interferon-{gamma} (IFN{gamma}) or interleukin-2 (IL-2), and xenograft models. However, the in vitro measurements did not reflect CAR-T cytotoxicity, whereas xenograft models are low throughput and costly. Here we presented a robust in vitro droplet microfluidic assay for CAR-T cytotoxicity assessment. This method not only enabled assessment of CAR-T cytotoxic activity under different fluid viscosity conditions, but also facilitated measurement of CAR-T expansion and dissection of mechanism of action via phenotype analysis in vitro. Furthermore, our data suggested that label-free cytotoxicity analysis is feasible by acquiring data before and after treatment. Hence, this study presented a novel in vitro method for assessment of cellular cytotoxicity that could potentially be applied to any cell-kill-cell experiment with varying solvent composition.

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Wong, K. U., Shi, J., Li, P., Wang, H., Jia, Y., Deng, C., Jiang, L., Wong, A. H.-H.. 2021-12-30. Assessment of chimeric antigen receptor T (CAR-T) cytotoxicity by droplet microfluidics in vitro. https://doi.org/10.1101/2021.12.28.474351

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