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CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios
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DOI：10.1039/D6RA02345B.png)
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Abstract
En 中文
Immune–cancer cell interactions play a central role in understanding antitumor responses and evaluating immunotherapies. However; long-term; single-cell-level analysis of these interactions remains challenging. To address this; we developed CellTrap; an instrument-free; perfusion-capable microfluidic device featuring 1024 parallel traps. Each trap is equipped with a filter constriction to stably retain cells under hydrostatic flow; sustain continuous medium perfusion; and minimize trap-to-trap crosstalk. By intentionally leveraging stochastic Poisson loading; a single seeding step simultaneously generates perfectly matched internal controls alongside variable effector-to-target (E : T) ratios across the array. Device characterization using 10 µm fluorescent beads and cells validated the predictable trap occupancy governed by Poisson statistics. Initial proof-of-concept experiments using primary human PBMCs against GFP-expressing U87 (U87GFP) glioblastoma cells successfully demonstrated targeted; immune-mediated cytotoxicity over 14 hours. To decouple donor-derived biological heterogeneity from technical validation; we subsequently employed IL-2-stimulated Natural Killer cells (NK92IL2) as a standardized effector population against U87GFP; K562 leukemia; and LS174T adenocarcinoma targets. Continuous time-lapse imaging seamlessly linked early transient intracellular calcium fluxes (indicating target engagement) to distinct; contact-dependent cytotoxic outcomes. Our data demonstrate that increasing E : T ratios consistently enhances immune-mediated target lysis; highlighting the platform's robust utility for dissecting complex immune–cancer dynamics and guiding the development of personalized immunotherapies.
Keywords:
CellTrap
microfluidic platform
immune–cancer interactions
effector-to-target ratios
single-cell analysis