CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios

🔗 원문 전체 보기 → Royal Society of Chemistry

요약

RSC Adv., 2026, 16,19024-19037DOI: 10.1039/D6RA02345B, Paper Open Access &nbsp This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.Muhammad Zia Ullah Khan, Morteza Hasanzadeh Kafshgari, Ali Bashiri Dezfouli, Oliver Hayden, Gabri…

본문

CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios

* Corresponding authors

a Control and Manipulation of Microscale Living Objects, Center for Translational Cancer Research (TranslaTUM), Munich Institute of Biomedical Engineering (MIBE), Munich Institute of Integrated Materials, Energy and Process Engineering (MEP), Department of Electrical Engineering, School of Computation, Information and Technology (CIT), Technical University of Munich (TUM), Einsteinstraße 25, Munich 81675, Germany
E-mail: [email protected]

b Heinz-Nixdorf-Chair of Biomedical Electronics, TranslaTUM, MIBE, School of Computation, Information and Technology, Technical University of Munich, Einsteinstraße 25, 81675 Munich, Germany

c Experimental Radiation Oncology and Radiobiology, TranslaTUM, School of Medicine, Technical University of Munich, Einsteinstraße 25, 81675 Munich, Germany

d Department of Otolaryngology, Head and Neck Surgery, TUM School of Medicine and Health, Technical University of Munich, 81675 Munich, Germany

Abstract

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.