StemXVivo Serum-Free Dendritic Cell Base Media
R&D Systems, part of Bio-Techne | Catalog # CCM003
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, CD14+ monocytes are differentiated into dendritic cells using the following procedure:
- Culture CD14+ cells in StemXVivo® Serum-Free Dendritic Cell Base Media supplemented with GM-CSF, IL-4, and Gentamicin.
- Replace the media every three days.
- Harvest immature MoDCs or use LPS to induce maturation
Reagents supplied in the StemXVivo® Serum-Free Dendritic Cell Base Media (Catalog # CCM003):
- 250 mL StemXVivo® Serum-Free Dendritic Cell Base Media
Reagents
- Human CD14+ monocytes isolated using the MagCellect Human CD14+ Cell Isolation Kit (Catalog # MAGH105) or equivalent
- Recombinant Human GM-CSF (Catalog # 215-GM)
- Recombinant Human IL-4 (Catalog # 204-IL)
- LPS
- Gentamicin (Catalog # B20192)
Materials
- Tissue culture flasks (25 cm2 or 75 cm2) or plates (6-well or 24-well)
- 15 mL centrifuge tubes
- Serological pipette
- Pipettes and pipette tips
- Hemocytometer
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge
- Vortex mixer
- Inverted Microscope
Differentiation of CD14+ Monocytes into Monocyte-derived Dendritic Cells Using StemXVivo® Serum-Free Dendritic Cell Base Media (Catalog # CCM003)
Thaw Human StemXVivo® Serum-Free Dendritic Cell Base Media.
Mix the thawed media by vortexing and warm the media in a 37 °C incubator.
Prepare the CD14+ cells from peripheral blood mononuclear cells.
Resuspend the purified CD14+ cells in Human StemXVivo® Serum-Free Dendritic Cell Base Media.
Perform a cell count and adjust the cell density to 1 x 106 cells/mL.
Supplement the media with Gentamicin, GM-CSF, and IL-4.
Add the cell suspension to the tissue culture flask or tissue culture plate.
Incubate the cells in a 37 °C and 5% CO2 humidified incubator for 7 days.
Change the media every three days.
Observe immature monocyte-derived dendritic cells (MoDCs) from day 5 to day 7.
Induce maturation of MoDCs by adding LPS to the cell suspension on day 7 and incubating for 48 hours.