CellXVivo Mouse Dendritic Cell Differentiation Kit
R&D Systems, part of Bio-Techne | Catalog # CDK008
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, immature and mature mouse dendritic cells can be generated from bone marrow using the following procedure:
- Isolate mouse bone marrow
- Culture bone marrow cells in growth factor-supplemented Differentiation Media
- Verify differentiation into immature dendritic cells by flow cytometry
- Induce dendritic cell maturation with TNF-alpha
- Verify dendritic cell maturation via flow cytometry
Reagents Supplied in the CellXVivo™ Mouse Dendritic Cell Differentiation Kit (Catalog # CDK008)
- Mouse Dendritic Cell Base Media
- Recombinant Mouse GM-CSF
- Recombinant Mouse IL-4
- Recombinant Mouse TNF-alpha
- Erythrocyte Lysing Buffer
- Reconstitution Buffer 1
Materials
- Laboratory mice
Equipment
- Syringe and needle
- Tissue culture flasks and/or plates
- Phosphate buffered saline (PBS)
- Sterile deionized water
- Microscope
- Hemocytometer
- 37 °C, 5% CO2 incubator
- Centrifuge
- Pipettes and pipette tips
Excise tibias and femurs from mouse.
Collect bone marrow cells.
Lyse red bloods cells with Erythrocyte Lysing Buffer.
Perform a cell count.
Suspend 1 x 106 bone marrow cells/mL in Differentiation Media.
Culture the cells for 5 days.
Add fresh Differentiation Media on day 3.
Verify immature dendritic cell differentiation by analyzing cell surface marker expression via flow cytometry (optional).
Immature dendritic cells are ready for desired application.
Induce dendritic cell maturation with TNF-α and incubate for 1 day.
Verify dendritic cell maturation by analyzing cell surface marker expression via flow cytometry
(optional).
Mature dendritic cells are ready for desired application.