Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro

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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-γ (IFNγ) 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 cytotoxicity experiment with varying solvent composition.
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Funding Information
  • Science and Technology Development Fund (137/2016/A, 110/2016/A3)
  • Chinese Academy of Sciences (XDB19030205)
  • Guangdong Provincial Applied Science and Technology Research and Development Program (2016B020237006)
  • Guangzhou Science and Technology Plan (201907010042, 201904010473)
  • Frontier Research Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory (2018GZR110105003)
  • UM Chair Professor (CPG2020-00004-FHS)

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