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Evaluation of Single-Cell Cytokine Secretion and Cell-Cell Interactions with a Hierarchical Loading Microwell Chip
Yufu Zhou1, Ning Shao2, Ricardo Bessa de Castro2
1Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030, USA; The Third Xiangya Hospital, Central South University, Changsha 410008, China; Center for inflammation and Epigenetics, Houston Methodist Research Institute, Houston, TX 77030, USA.
Insights
A novel hierarchical loading microwell chip (HL-Chip) enables precise, high-throughput evaluation of individual T cell functions, including cytokine secretion and cytolysis, crucial for advancing adoptive cell therapy (ACT). This technology offers a robust platform for detailed single-cell analysis.
Area of Science:
- Biotechnology
- Immunology
- Cell Biology
Background:
- Current methods struggle to comprehensively assess individual T cell functions like cytokine secretion and cytolysis.
- Accurate single-cell analysis is critical for optimizing adoptive cell therapy (ACT) efficacy and safety.
Purpose of the Study:
- To develop and validate a novel microwell chip technology for high-throughput, single-cell functional analysis.
- To demonstrate the utility of this chip in evaluating T cell secretory profiles, cytolytic activity, and tumor cell interactions.
Main Methods:
- Development of a hierarchical loading microwell chip (HL-Chip) for precise alignment of single cells and functionalized beads.
- Utilizing size-based sorting within microwells for high-throughput single-cell/bead combinations.
- Application of the HL-Chip for longitudinal secretory profiling, cytolytic activity assessment, and integrated cell-cell interaction studies.
Main Results:
- The HL-Chip successfully achieved single-cell/bead precision in high-throughput microwell arrays.
- Demonstrated capability for longitudinal secretory profiling of individual T cells.
- Enabled large-scale evaluation of single T cell cytolytic activity and T cell-tumor cell interactions.
Conclusions:
- The HL-Chip is a simple, robust, and versatile technology for comprehensive single T cell functional evaluation.
- This platform significantly advances the ability to analyze T cell responses in detail.
- Facilitates improved understanding and development of adoptive cell therapies.
Abstract:
Comprehensive evaluation of single T cell functions such as cytokine secretion and cytolysis of target cells is greatly needed in adoptive cell therapy (ACT) but has never been fully fulfilled by current approaches. Herein, we develop a hierarchical loading microwell chip (HL-Chip) that aligns multiple cells and functionalized beads in a high-throughput microwell array with single-cell/bead precision based on size differences. We demonstrate the potential of the HL-Chip in evaluating single T cell functions by three applications: high-throughput longitudinal secretory profiling of single T cells, large-scale evaluation of cytolytic activity of single T cells, and integrated T cell-tumor cell interactions. The HL-Chip is a simple and robust technology that constructs arrays of defined cell/object combinations for multiple measurements and material retrieval.
