A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns

Abstract

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.

AI-Powered Microfluidics: Shaping the Future of Phenotypic Drug Discovery

“…Moreover, coculturing different cell types on microfluidic devices such as cancer cells and immune cells allows for the study of complex cell–cell interactions that are important in the development of cancer immunotherapies. Cui et al introduced a versatile microfluidic immunoassay that integrates microtopographic cell-culture substrates with a microbeads-based immunofluorescence assay for rapid and sensitive analysis . Parlato et al reported a microfluidic platform composed of five parallel channels capable of assessing the guided efficient motion of Interferon-α-conditioned dendritic cells toward cancer cells and subsequent phagocytosis .…”

“…Cui et al introduced a versatile microfluidic immunoassay that integrates microtopographic cell-culture substrates with a microbeads-based immunofluorescence assay for rapid and sensitive analysis. 26 Parlato et al reported a microfluidic platform composed of five parallel channels capable of assessing the guided efficient motion of Interferon-αconditioned dendritic cells toward cancer cells and subsequent phagocytosis. 27 Tu et al developed a microfluidic device that allows for comprehensive analysis of immunocyte heterogeneity during interactions with tumor cells at the single-cell level, uncovering the effects of immune checkpoint blockade in cancer.…”

Quantitatively Evaluating Interactions between Patient-Derived Organoids and Autologous Immune Cells by Microfluidic Chip

“…29 Apart from evaluating the cytokine secreted by a single type of cell, studies for evaluating interactions between multiple types of cells were also presented. 30 The antibody-assisted enrichment 31 and immunofluorescence 32 to detect cytokines were also proven feasible in the scenario of coculturing multiple kinds of cells in microfluidic devices.…”

Characterization of Shrink Film Properties for Rapid Microfluidics Lab-on-Chip Fabrication

“…Microfluidics involves the examination and control of minute quantities of fluids, typically in the microliter or nanoliter range. This field has gained significant prominence in research due to its diverse applications in areas like biotechnology, pharmaceuticals, diagnostics, lab-on-chip (LOC), separation and isolation of circulating tumor cells, sperm sorting, and bacteria or algae detection [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 ]. The integration of microscale structures on a single chip has revolutionized research across various domains by enabling the swift manipulation, analysis, and distribution of small samples.…”