Analyzing functional heterogeneity of effector cells for enhanced adoptive cell therapy applications

Area of Science:

• Biotechnology
• Immunology
• Cell Biology

Background:

• Traditional cellular effector function assays use bulk populations, obscuring cellular heterogeneity and rare cell subsets.
• Understanding individual cell behavior is crucial for optimizing cellular therapies.

Purpose of the Study:

• To introduce and validate Xdrop® double-emulsion droplet technology for high-throughput single-cell analysis.
• To assess the utility of this technology for studying cellular effector functions in various cell types relevant to adoptive cell therapy.

Main Methods:

• Utilized Xdrop® droplet technology for compartmentalizing single cells or cell pairs in double emulsions.
• Evaluated effector cell molecule secretion and target cell killing at the single-cell level.
• Demonstrated compatibility with standard laboratory techniques like flow cytometry and microscopy for data acquisition.
• Showcased viable single-cell isolation for further analysis or expansion.

Main Results:

• Successfully applied double-emulsion droplet technology to natural killer cells, T cells, tumor-infiltrating lymphocytes, and chimeric antigen receptor T cells.
• Achieved high-throughput compartmentalization and single-cell level readouts of cellular functions.
• Confirmed the ability to independently or simultaneously assess molecule secretion and cell killing.

Conclusions:

• Xdrop® droplet technology provides unparalleled resolution for single-cell analysis of cellular effector functions.
• This technology is a valuable tool for advancing the development and understanding of cellular therapy products.
• Enables detailed characterization of cellular heterogeneity and rare subpopulations in therapeutic cell populations.