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Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses
Published on: March 8, 2024
On-chip immune cell activation and subsequent time-resolved magnetic bead-based cytokine detection
Patthara Kongsuphol1, Yunxiao Liu2, Qasem Ramadan2
1Institute of Microelectronics, Agency for Science Technology and Research (A*STAR), 2 Fusionopolis Way, #08-02 Innovis Tower, Singapore, 138634, Singapore. kongsupholp@ime.a-star.edu.sg.
Insights
This study presents a novel microfluidic device for real-time cytokine detection from single cell clusters. The in situ magnetic immune assay enables precise monitoring of immune responses for disease research and personalized medicine.
Area of Science:
- Biotechnology
- Immunology
- Microfluidics
Background:
- Cytokine profiling and immunophenotyping are crucial for understanding disease mechanisms, personalized diagnosis, and immunotherapy.
- Current methods may lack the temporal resolution or cellular specificity needed for detailed analysis.
Purpose of the Study:
- To develop a time-resolved, in situ method for detecting cytokines secreted by single cell clusters.
- To enable simultaneous cell culture under perfusion and cytokine detection at multiple time-points.
Main Methods:
- Fabrication of a triple-layered microfluidic chamber with separate compartments for cell culture, perfusion, and immunoassay.
- Utilizing porous membranes for nutrient diffusion and cytokine capture without disturbing cell culture.
- Employing functionalized magnetic beads for in situ cytokine capturing and quantification.
Main Results:
- Demonstrated successful time-resolved detection of cytokines from single cell clusters.
- Quantified cytokines released from U937 monocytes and differentiated macrophages under differential stimuli.
- The microfluidic design facilitated cytokine capture while maintaining cell culture integrity and perfusion benefits.
Conclusions:
- The developed in situ magnetic immune assay offers a powerful tool for real-time cytokine analysis in cellular microenvironments.
- This technology holds promise for advancing research in disease mechanisms, diagnostics, and the development of immunotherapies.
Abstract:
Cytokine profiling and immunophenotyping offer great potential for understanding many disease mechanisms, personalized diagnosis, and immunotherapy. Here, we demonstrate a time-resolved detection of cytokine from a single cell cluster using an in situ magnetic immune assay. An array of triple-layered microfluidic chambers was fabricated to enable simultaneous cell culture under perfusion flow and detection of the induced cytokines at multiple time-points. Each culture chamber comprises three fluidic compartments which are dedicated to, cell culture, perfusion and immunoassay. The three compartments are separated by porous membranes, which allow the diffusion of fresh nutrient from the perfusion compartment into the cell culture compartment and cytokines secretion from the cell culture compartment into the immune assay compartment. This structure hence enables capturing the released cytokines without disturbing the cell culture and without minimizing benefit gain from perfusion. Functionalized magnetic beads were used as a solid phase carrier for cytokine capturing and quantification. The cytokines released from differential stimuli were quantified in situ in non-differentiated U937 monocytes and differentiated macrophages.
