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Isolation and generation of human dendritic cells
Smita Nair1, Gerald E Archer1, Thomas F Tedder1
1Duke University Medical Center, Durham, North Carolina.
Current Protocols in Immunology
|November 7, 2012
Summary
This study details methods to isolate and generate dendritic cells (DCs), crucial antigen-presenting cells, from human blood. Researchers can enrich DC populations or generate them from monocytes for various immunological studies.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are specialized antigen-presenting cells (APCs) vital for initiating immune responses.
- Mature DCs typically constitute a small fraction (∼0.2%) of human blood mononuclear cells.
Purpose of the Study:
- To describe methods for isolating and generating dendritic cells from human blood.
- To provide techniques for enriching DC populations and depleting them for comparative studies.
Main Methods:
- Cell enrichment techniques to increase the frequency of mature blood dendritic cells.
- Fluorescence-activated cell sorting for highly purified dendritic cell preparations.
- Generation of dendritic cells from monocytes using cytokine culture.
- Negative selection methods to deplete dendritic cells for control experiments.
Main Results:
- Established protocols allow for significantly increased yields of dendritic cells from human blood.
- Purified dendritic cell populations can be obtained through cell sorting.
- Dendritic cells can be effectively generated in vitro from monocyte precursors.
- Methods for dendritic cell depletion are available for functional comparisons.
Conclusions:
- The described methods provide versatile approaches for obtaining and manipulating dendritic cells for immunological research.
- These techniques enhance the ability to study dendritic cell function and their role in immune responses.
