Related Experiment Video
Updated: Nov 20, 2025
Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
Purification of Murine IL-10 + B Cells for Analyses of Biological Functions and Transcriptomics
1Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, China. sunjb3@mail.sysu.edu.cn.
Insights
Regulatory B cells (Breg) play a crucial role in immune suppression. CD9 is identified as a key marker for B10 cells in mice, aiding in their isolation and study.
Area of Science:
- Immunology
- Cell Biology
Background:
- B10 cells are a subset of regulatory B cells (Breg) known for immune suppression via IL-10.
- Identifying and isolating B10 cells is critical for understanding their function.
- CD9 has been recently identified as a specific marker for murine B10 cells.
Purpose of the Study:
- To provide standardized methods for the characterization and functional evaluation of CD9+ B cells.
- To enable the purification and subsequent analysis of B10 cells.
Main Methods:
- Preparation of leukocytes.
- Characterization of CD9+ B cells using multiple assays.
- Evaluation of CD9+ B-cell activity.
- RNA purification from B10 cells for transcriptomic analysis.
Main Results:
- Established five distinct methods for B10 cell analysis.
- Demonstrated the utility of CD9 in distinguishing B10 cells from other B cell populations.
- Enabled detailed molecular analysis through transcriptomics.
Conclusions:
- CD9 is a reliable marker for identifying and purifying murine B10 cells.
- The provided methods facilitate comprehensive study of B10 cell biology and function.
- This work supports further research into the immunomodulatory roles of B10 cells.
Abstract:
B10 cells are the most frequently investigated subset of Breg cells, capable of suppressing immunity through the expression of the immunosuppressive cytokine IL-10. B10 cells are enriched in phenotypically diverse B-cell subsets. Recently, CD9 was identified as a marker of B10 cells in mice (human B10 cells have a separate set of markers that do not overlap with murine B10 cells). Together with a combination of other B10 markers, CD9 can be used to distinguish both mature and immature B10 cells from nonregulatory B cells and support selective purification of B10 cells. Here we provide five methods for the characterization and activity evaluation of CD9+ B cells. The first method is used for the preparation of leukocytes, the second and third are used for the characterization of CD9+ B cells, while the last two methods serve to evaluate CD9+ B-cell activities. Finally, we detail the purification of RNA from B10 cells and the performance of transcriptomic assays.
