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Protocol for the isolation and purification of human follicular dendritic cells for functional assays
Martijn Breeuwsma1, Balthasar A Heesters1
1Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, the Netherlands.
Insights
This protocol details isolating human follicular dendritic cells (FDCs) from lymphoid tissues. This method aids in studying FDC roles in antibody development and B cell interactions.
Area of Science:
- Immunology
- Cell Biology
Background:
- Follicular dendritic cells (FDCs) are crucial for adaptive immunity.
- FDCs present antigens to B cells within germinal centers, essential for antibody maturation.
- Dysfunction in FDC activity is linked to various immune disorders.
Purpose of the Study:
- To provide a detailed protocol for isolating and purifying human follicular dendritic cells (FDCs).
- To enable downstream functional and descriptive assays on isolated FDCs.
- To establish a reliable method applicable to diverse human lymphoid tissues.
Main Methods:
- Enzymatic digestion of lymphoid tissues.
- Fluorescence-activated cell sorting (FACS) for FDC purification.
- Protocol validated on tonsils, lymph nodes, and tertiary lymphoid structures.
Main Results:
- Successful isolation and purification of viable human FDCs.
- Demonstrated applicability across multiple human lymphoid tissue types.
- Established a robust method for obtaining FDCs for further research.
Conclusions:
- The described protocol offers a reliable method for human FDC isolation.
- This technique supports advanced studies on FDC function in B cell immunity.
- Facilitates research into immune responses and related pathologies.
Abstract:
In this protocol, we detail how to isolate and purify human follicular dendritic cells (FDCs) from lymphoid tissues. FDCs play a vital role in antibody development by presenting antigens to B cells in germinal centers. The assay involves enzymatic digestion and fluorescence-activated cell sorting and is successfully applied to various lymphoid tissues, including tonsils, lymph nodes, and tertiary lymphoid structures. Our robust technique enables the isolation of FDCs and facilitates downstream functional and descriptive assays. For complete details on the use and execution of this protocol, please refer to Heesters et al.1.
