Xie Yuanyuan, Ning Ke, Sun Wen, Feng Lingke, Chen Yirong, Sun Wei, Li Yan, Yu Ling
Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing 400715, China.
Key Laboratory of Animal Biological Products & Genetic Engineering, Ministry of Agriculture and Rural, Sinopharm Animal Health Corporation Ltd., Wuhan 430023, China; State Key Laboratory of Novel Vaccines for Emerging Infectious Diseases, China National Biotec Group Company Limited, Beijing 100024, China.
J Biotechnol. 2025 Jan;397:11-21. doi: 10.1016/j.jbiotec.2024.11.008. Epub 2024 Nov 15.
Natural killer (NK) cells are pivotal in immunotherapy due to their potent tumor-targeting capabilities. However, accessible in vitro 3D dynamic models for evaluating Tumor Infiltrating Natural Killer Cells (TINKs) remain scarce. This study addresses this gap by developing a novel pump-free microfluidic chip to investigate the interactions between NK-92 cells and prostate DU 145 tumor spheroids. The platform facilitates the separation of free NKs and TINKs for subtype characterization. The design integrates multiple planes with a multi-layer paper scaffold to accommodate tumor spheroids, allowing NK-92 cells to traverse Matrigel-coated barriers that mimic the extracellular matrix. The dual-channel pump-free device enables unidirectional circulation of NK-92 cells, allowing analysis of tumor spheroid movement and NK-92 cell interactions under flow conditions. Results demonstrate continuous fluid circulation in the dual-channel device by rocking the platform at tilt angles of 21° and 15°. Tumor spheroids show- enhanced migration under flow conditions compared to static culture. Although spheroids recruit slightly more NK-92 cells under flow conditions, CD56 and CD16 receptor expression on IL-2-activated free NK-92 cells and tumor-infiltrating NK-92 cells matches in vivo patterns in dynamic cultures. These findings suggest that tumor cells and fluid dynamics significantly influence NK cell subtypes. This pump-free microfluidic platform is a functional tool for simulating and studying immune cell-tumor interactions, providing valuable insights into NK cell dynamics with tumor spheroids in physiologically relevant environments.
自然杀伤（NK）细胞因其强大的肿瘤靶向能力在免疫治疗中起着关键作用。然而，用于评估肿瘤浸润自然杀伤细胞（TINKs）的可及的体外3D动态模型仍然稀缺。本研究通过开发一种新型无泵微流控芯片来填补这一空白，以研究NK-92细胞与前列腺DU 145肿瘤球体之间的相互作用。该平台有助于分离游离的NK细胞和TINKs以进行亚型表征。该设计将多个平面与多层纸质支架整合在一起以容纳肿瘤球体，使NK-92细胞能够穿过模拟细胞外基质的基质胶包被的屏障。这种双通道无泵装置能够使NK-92细胞单向循环，从而能够在流动条件下分析肿瘤球体的运动和NK-92细胞的相互作用。结果表明，通过以21°和15°的倾斜角度摇晃平台，双通道装置中可实现连续的流体循环。与静态培养相比，肿瘤球体在流动条件下显示出增强的迁移。尽管在流动条件下球体募集的NK-92细胞略多，但IL-2激活的游离NK-92细胞和肿瘤浸润NK-92细胞上的CD56和CD16受体表达与动态培养中的体内模式相匹配。这些发现表明肿瘤细胞和流体动力学显著影响NK细胞亚型。这种无泵微流控平台是模拟和研究免疫细胞与肿瘤相互作用的功能性工具，为在生理相关环境中研究NK细胞与肿瘤球体的动态关系提供了有价值的见解。
