Droplet Microfluidics Enables Tracing of Target Cells at the Single-Cell Transcriptome Resolution.

Area of Science:

• Biotechnology
• Genomics
• Immunotherapy

Background:

• Single-cell omics techniques are advancing precision medicine, prenatal diagnosis, and embryo development.
• Existing single-cell workflows require multiple steps like demulsification and cell recovery, impacting efficiency.
• There is a growing demand for updated single-cell techniques, especially for specific target cells.

Purpose of the Study:

• To develop an integrated, labor-saving method for monitoring TCR-T cells using single-cell transcriptome profiling.
• To streamline the workflow from cell encapsulation to single-cell sequencing within droplets.
• To reduce operational complexity and improve the efficiency of T cell analysis for immunotherapy.

Main Methods:

• An all-in-droplet method was developed, integrating cell encapsulation, target sorting, droplet picoinjection, and single-cell transcriptome profiling on chips.
• TCR-T cells were encapsulated and sorted within droplets, followed by reagent injection for scRNA-seq.
• The method avoided droplet breakage and re-encapsulation steps between sorting and sequencing.

Main Results:

• Achieved excellent recovery of single-cell transcriptome data with a median gene number exceeding 4000.
• Reported a low cross-contamination rate of 8.2 ± 2%.
• Demonstrated a convenient and highly integrable device for monitoring TCR-T cells.

Conclusions:

• The developed all-in-droplet method significantly simplifies TCR-T cell monitoring and single-cell transcriptome sequencing.
• This labor-saving strategy accelerates the progress of chip-based cell analysis and marketization.
• The technology holds promise for advancing adoptive T cell immunotherapy and its clinical applications.