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A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns
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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687854/ https://www.ncbi.nlm.nih.gov/pubmed/36354472 http://dx.doi.org/10.3390/bios12110963 |
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author | Cui, Xin Liu, Lelin Li, Jiyu Liu, Yi Liu, Ya Hu, Dinglong Zhang, Ruolin Huang, Siping Jiang, Zhongning Wang, Yuchao Qu, Yun Pang, Stella W. Lam, Raymond H. W. |
author_facet | Cui, Xin Liu, Lelin Li, Jiyu Liu, Yi Liu, Ya Hu, Dinglong Zhang, Ruolin Huang, Siping Jiang, Zhongning Wang, Yuchao Qu, Yun Pang, Stella W. Lam, Raymond H. W. |
author_sort | Cui, Xin |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9687854 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96878542022-11-25 A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns Cui, Xin Liu, Lelin Li, Jiyu Liu, Yi Liu, Ya Hu, Dinglong Zhang, Ruolin Huang, Siping Jiang, Zhongning Wang, Yuchao Qu, Yun Pang, Stella W. Lam, Raymond H. W. Biosensors (Basel) Article 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. MDPI 2022-11-02 /pmc/articles/PMC9687854/ /pubmed/36354472 http://dx.doi.org/10.3390/bios12110963 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Cui, Xin Liu, Lelin Li, Jiyu Liu, Yi Liu, Ya Hu, Dinglong Zhang, Ruolin Huang, Siping Jiang, Zhongning Wang, Yuchao Qu, Yun Pang, Stella W. Lam, Raymond H. W. A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title | A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title_full | A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title_fullStr | A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title_full_unstemmed | A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title_short | A Microfluidic Platform Revealing Interactions between Leukocytes and Cancer Cells on Topographic Micropatterns |
title_sort | microfluidic platform revealing interactions between leukocytes and cancer cells on topographic micropatterns |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687854/ https://www.ncbi.nlm.nih.gov/pubmed/36354472 http://dx.doi.org/10.3390/bios12110963 |
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