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IL-2 secretion-based sorting of single T cells using high-throughput microfluidic on-cell cytokine capture
Robert Dimatteo1,2, Dino Di Carlo2,3,4
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, 5531 Boelter Hall, P.O. Box 951592, Los Angeles, CA, 90095, USA.
Lab on a Chip
|March 16, 2022
Summary
Researchers developed a new workflow to analyze and sort T cells by their IL-2 secretion using droplets. This method improves accuracy and accessibility for immune cell analysis in therapies.
Area of Science:
- Immunology
- Biotechnology
- Cell Biology
Background:
- Secreted proteins are vital for immune responses, particularly in engineered T cell therapies.
- Current methods for analyzing and sorting cells by secretion are limited in accessibility and accuracy.
Purpose of the Study:
- To develop a rapid, accessible workflow for screening and sorting single T cells based on their IL-2 secretion.
- To overcome limitations of existing methods in analyzing cellular secretory functions.
Main Methods:
- Utilized nanoliter droplets for cell partitioning, enabling IL-2 accumulation on cell surfaces without crosstalk.
- Employed step emulsification for monodisperse droplet generation at high flow rates.
- Disrupted emulsions for downstream analysis using conventional flow cytometry and fluorescence-activated cell sorting (FACS).
Main Results:
- Achieved high-throughput processing (200 μL min⁻¹) with minimal secretion crosstalk.
- Demonstrated a 10-fold increase in dynamic range for IL-2 detection compared to unencapsulated cells.
- Successfully sorted IL-2 secreting T cells using FACS, showing no crosstalk after three hours.
Conclusions:
- The developed workflow enables accurate and accessible sorting of T cells based on functional secretory properties.
- This method significantly enhances the analysis of immune cell function for applications like T cell therapy.
- The approach offers a more precise and scalable solution for studying cellular secretions.
