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Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging
Published on: July 9, 2008
Generation of dendritic cells from rat bone marrow.
T Powell1, J R Major, G Macpherson
1Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
Methods in Molecular Medicine
|March 5, 2011
Summary
Generating dendritic cells (DC) from bone marrow in vitro is now feasible using recombinant cytokines like granulocyte-macrophage colony-stimulating factor (GM-CSF). This method yields functional DC for research and potential therapeutic applications.
Area of Science:
- Immunology
- Cell Biology
- Hematopoiesis
Background:
- Dendritic cells (DC) are crucial immune cells derived from bone marrow.
- Previous in vitro methods yielded limited DC numbers due to insufficient maturation and survival signals.
- Advances in cytokine availability revolutionized DC research.
Purpose of the Study:
- To describe a method for generating functional dendritic cells (DC) from murine bone marrow in vitro.
- To overcome limitations of previous DC generation techniques.
Main Methods:
- Depletion of erythrocytes, B cells, and T cells from murine bone marrow.
- Culture of depleted bone marrow cells with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF).
- Assessment of generated cells via phenotype, function, and allogeneic mixed lymphocyte reaction (MLR).
Main Results:
- Culturing bone marrow cells with GM-CSF for 7 days produced typical DC.
- Generated DC exhibited stimulatory capacity in the allogeneic MLR, a key indicator of DC function.
- These DC expressed high levels of MHC class I, MHC class II, and costimulatory molecules.
Conclusions:
- Recombinant GM-CSF enables efficient in vitro generation of functional dendritic cells from murine bone marrow.
- This method provides a valuable tool for DC research and potential clinical applications.
- Human DC culture remains challenging due to difficulties in obtaining sufficient cell numbers.
