Park, Junhyun Kim, Seong-Eun Kim, Jaejeung Yoon, Minjung Doh, Junsang Hyun, Kyung-A Jung, Hyo-Il 2024-09-14T06:00:27Z 2024-09-14T06:00:27Z 2024-09-13 2025-02 2365-709X https://pubs.kist.re.kr/handle/201004/150575 Circulating tumor cell (CTC) clusters represent formidable precursors of cancer metastasis due to their heightened immune resistance against natural killer (NK) cells. Despite this, the cytotoxicity of NK cells against CTC clusters, particularly their interaction with other immune cells such as neutrophils, remains inadequately examined. This study introduces a dual-nozzle integrated droplet microfluidic chip (dual-nozzle chip) designed to facilitate the deterministic encapsulation of three distinct cell types-CTCs, NK cells, and neutrophils-to monitor the dynamic cytotoxicity between immune cells and target cells. The dual-nozzle chip comprises double-spiral channels and a serpentine channel for inertial cell focusing, alongside dual-nozzle oil phases employed to generate monodisperse droplets at high flow rates. Utilizing Rayleigh-Plateau instability, the focused cell streams, characterized by high inertia, undergo pinching off into monodisperse droplets at the flow-focusing junction, where dual-nozzle oil phases are introduced. Consequently, triple cells are paired at the desired ratios, overcoming the intrinsic challenge posed by the Poisson distribution. A droplet-based assay demonstrates that NK cell-mediated cytotoxicity varies depending on the type of cancer cells and the presence of suppressor cells. The design strategy of the dual-nozzle chip exhibits promises for broader applications, emphasizing its potential for analyzing diverse cell-to-cell interactions. This study introduces a dual-nozzle integrated droplet microfluidic chip designed for screening the cytotoxicity of three cell types. The chip features inertial focusing channels, double-spiral and serpentine channels, and the dual-nozzle flow-focusing junction for monodisperse droplet generation, thereby achieving deterministic encapsulation and cell pairing. It demonstrates that NK cell-mediated cytotoxicity varies based on the cancer cell form and neutrophil incorporation. image English JOHN WILEY & SONS INC Dual Nozzle-Assisted Deterministic Encapsulation of Triple Particles for Screening NK-Cell Cytotoxicity Against Circulating Tumor Cell Clusters Article 10.1002/admt.202400477 1 Advanced Materials Technologies, v.10, no.3 Advanced Materials Technologies 10 3 N scie scopus 001303074800001 2-s2.0-85202693666 Materials Science, Multidisciplinary Materials Science Article MIGRATION SYSTEM NATURAL-KILLER-CELLS DROPLET FORMATION LABEL-FREE FLOW Rayleigh-Plateau instability circulating tumor cell clusters droplet generation dual-nozzle NK cell cytotoxicity