Department of Biomedical Engineering, School of Medical Technology, Beijing Institute of Technology, Beijing 100081, China.
Department of Thoracic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China.
Anal Chem. 2024 Aug 13;96(32):13061-13069. doi: 10.1021/acs.analchem.4c01389. Epub 2024 Aug 2.
The coculture of patient-derived tumor organoids (PDOs) and autologous immune cells has been considered as a useful surrogate of tumor-immune environment. However, the immune interactions between PDOs and autologous immune cells, including immune-mediated killing behaviors and immune-related cytokine variations, have yet to be quantitatively evaluated. This study presents a microfluidic chip for quantifying interactions between PDOs and autologous immune cells (IOI-Chip). A baffle-well structure is designed to ensure efficient trapping, long-term coculturing, and fluorescent observation of a limited amount of precious PDOS and autologous immune cells, while a microbeads-based immunofluorescence assay is designed to simultaneously quantify multiple kinds of immune-related cytokines . The PDO apoptosis and 2 main immune-related cytokines, TNF-α and IFN-γ, are simultaneously quantified using samples from a lung cancer patient. This study provides, for the first time, a capability to quantify interactions between PDOs and autologous immune cells at 2 levels, the immune-mediated killing behavior, and multiple immune-related cytokines, laying the technical foundation of assessment of patient immune response.
患者来源的肿瘤类器官（PDO）与自体免疫细胞的共培养被认为是一种有用的肿瘤免疫环境替代物。然而，PDO 与自体免疫细胞之间的免疫相互作用，包括免疫介导的杀伤行为和免疫相关细胞因子的变化，尚未进行定量评估。本研究提出了一种用于定量分析 PDO 与自体免疫细胞相互作用的微流控芯片（IOI-Chip）。设计了一个挡板-井结构，以确保有限数量的珍贵 PDO 和自体免疫细胞的高效捕获、长期共培养和荧光观察，同时设计了基于微珠的免疫荧光分析，以同时定量多种免疫相关细胞因子。利用来自肺癌患者的样本，同时定量分析了 PDO 凋亡和 2 种主要的免疫相关细胞因子 TNF-α和 IFN-γ。本研究首次提供了定量分析 PDO 与自体免疫细胞相互作用的能力，包括免疫介导的杀伤行为和多种免疫相关细胞因子，为评估患者免疫反应奠定了技术基础。
