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Updated: Jun 12, 2025

Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

Published on: January 10, 2019

Protocol for mapping T cell activation using single-cell RNA-seq.

Hui Li¹, Yifei Liu², Xuefei Wang¹

¹CAS Key Laboratory of Computational Biology, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China; School of Life Sciences, Southern University of Science and Technology, Shenzhen 518055, China.

|October 20, 2024

Summary

This summary is machine-generated.

This study details a protocol for analyzing CD4 T-cell activation using anti-CD3/CD28 bead stimulation and single-cell RNA sequencing. The method covers cell isolation, stimulation, and data analysis for T-cell activation research.

Keywords:

Cell Biology Cell isolation Immunology Molecular Biology RNAseq Sequence analysis Sequencing Single Cell

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Area of Science:

Immunology
Molecular Biology
Genomics

Background:

CD4 T-cell activation is crucial in immune responses.
Anti-CD3/CD28 stimulation is a standard model for T-cell activation studies.
Single-cell RNA sequencing (scRNA-seq) offers high-resolution insights into cellular heterogeneity.

Purpose of the Study:

To present a comprehensive protocol for investigating T-cell activation.
To combine anti-CD3/CD28 bead stimulation with scRNA-seq for detailed analysis.
To provide a reproducible workflow for T-cell activation studies.

Main Methods:

Isolation of human peripheral blood mononuclear cells (PBMCs).
Enrichment of CD4 T-cells.
Stimulation using anti-CD3/CD28 beads.
Single-cell RNA sequencing (scRNA-seq) and data analysis.

Main Results:

A detailed workflow for T-cell activation analysis using scRNA-seq is established.
The protocol enables the study of T-cell responses at a single-cell level.
The method facilitates the understanding of molecular changes during T-cell activation.

Conclusions:

The presented protocol effectively integrates anti-CD3/CD28 stimulation with scRNA-seq.
This method provides a powerful tool for dissecting T-cell activation mechanisms.
The workflow supports advancements in immunological research and therapeutic development.