Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou 519070, China.
Department of Biomedical Engineering, City University of Hong Kong, Hong Kong 999077, China.
Biosensors (Basel). 2022 Nov 2;12(11):963. doi: 10.3390/bios12110963.
Immunoassay for detailed analysis of immune-cancer intercellular interactions can achieve more promising diagnosis and treatment strategies for cancers including nasopharyngeal cancer (NPC). In this study, we report a microfluidic live-cell immunoassay integrated with a microtopographic environment to meet the rising demand for monitoring intercellular interactions in different tumor microenvironments. The developed assay allows: (1) coculture of immune cells and cancer cells on tunable (flat or micrograting) substrates, (2) simultaneous detection of different cytokines in a wide working range of 5-5000 pg/mL, and (3) investigation of migration behaviors of mono- and co-cultured cells on flat/grating platforms for revealing the topography-induced intercellular and cytokine responses. Cytokine monitoring was achieved on-chip by implementing a sensitive and selective microbead-based sandwich assay with an antibody on microbeads, target cytokines, and the matching fluorescent-conjugated detection antibody in an array of active peristaltic mixer-assisted cytokine detection microchambers. Moreover, this immunoassay requires a low sample volume down to 0.5 μL and short assay time (30 min) for on-chip cytokine quantifications. We validated the biocompatibility of the co-culture strategy between immune cells and NPC cells and compared the different immunological states of undifferentiated THP-1 monocytic cells or PMA-differentiated THP-1 macrophages co-culturing with NP460 and NPC43 on topographical and planar substrates, respectively. Hence, the integrated microfluidic platform provides an efficient, broad-range and precise on-chip cytokine detection approach, eliminates the manual sampling procedures and allows on-chip continuous cytokine monitoring without perturbing intercellular microenvironments on different topographical ECM substrates, which has the potential of providing clinical significance in early immune diagnosis, personalized immunotherapy, and precision medicine.
免疫分析用于详细分析免疫-癌症细胞间相互作用，可以为包括鼻咽癌（NPC）在内的癌症实现更有前景的诊断和治疗策略。在这项研究中，我们报告了一种微流控活细胞免疫分析与微形貌环境相结合的方法，以满足监测不同肿瘤微环境中细胞间相互作用的需求。所开发的分析方法允许：（1）在可调（平面或微光栅）基底上共培养免疫细胞和癌细胞，（2）在 5-5000 pg/mL 的宽工作范围内同时检测不同的细胞因子，（3）在平面/光栅平台上研究单培养和共培养细胞的迁移行为，以揭示形貌诱导的细胞间和细胞因子反应。通过在微流控芯片上实施基于微珠的敏感和选择性夹心免疫分析，在一系列主动蠕动混合器辅助的细胞因子检测微室阵列中使用微珠上的抗体、靶细胞因子和匹配的荧光标记检测抗体，实现了细胞因子的芯片上监测。此外，这种免疫分析需要低至 0.5 μL 的样品体积和 30 分钟的短分析时间，用于在芯片上进行细胞因子定量。我们验证了免疫细胞与 NPC 细胞共培养策略的生物相容性，并比较了未分化的 THP-1 单核细胞或 PMA 分化的 THP-1 巨噬细胞与 NP460 和 NPC43 在形貌和平面基底上共培养的不同免疫状态。因此，集成的微流控平台提供了一种高效、宽范围和精确的芯片上细胞因子检测方法，消除了手动采样程序，并允许在不同形貌 ECM 基底上的芯片上连续细胞因子监测，而不会干扰细胞间微环境，这有可能在早期免疫诊断、个性化免疫治疗和精准医学中提供临床意义。
Zotero 插件
