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bioRxiv · 10.1101/2021.12.28.474355

In vitro cytokine expression analysis by droplet microfluidics

Wong, A. H.-H.·Akincilar, S. C.·Chua, J. Y. H.·Chanthira Morgan, D. d. K.·Hei, D.·Tergaonkar, V.
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Abstract

Droplet microfluidics provides a miniaturized platform to conduct biological assays. We previously developed a droplet microfluidic chip assay for screening cancer cells against chemical drugs and chimeric antigen receptor T (CAR-T) cells, respectively. In this study, we investigated chip application on a cytokine expression assay using MCF7 breast cancer reporter cells engineered by fusing green fluorescent protein (GFP) to the C-terminus of endogenous interleukin-6 (IL6) gene. Combined tumor necrosis factor (TNF) treatment and serum-free medium starvation stimulated IL6-GFP expression and enhanced GFP fluorescence. Our data showed that on-chip assay recapitulates the cellular response in vitro, although absolute quantification of IL6 induction could not be accomplished. The demonstration of multi-timepoint IL6 expression analysis paves the way for our future study on tumor response to immune attack via cytokine signaling.

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Wong, A. H.-H., Akincilar, S. C., Chua, J. Y. H., Chanthira Morgan, D. d. K., Hei, D., Tergaonkar, V.. 2021-12-28. In vitro cytokine expression analysis by droplet microfluidics. https://doi.org/10.1101/2021.12.28.474355

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