StemXVivo Serum-Free Dendritic Cell Base Media
R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
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
- 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)
- Gentamicin (Catalog # B20192)
- 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
- 37 °C and 5% CO2 incubator
- 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.
Citations for StemXVivo Serum-Free Dendritic Cell Base Media (2)
Citations are publications that use Bio-Techne products. Selected citations for StemXVivo Serum-Free Dendritic Cell Base Media include:
Sample Types: Whole Cells
Legat A et al. (2010-01-01), CD14-independent responses induced by a synthetic lipid A mimetic. Eur J Immunol, 40(3):797-802.
Sample Types: Whole Cells
Duraisingham SS et al. (2009-01-01), TLR-stimulated CD34 stem cell-derived human skin-like and monocyte-derived dendritic cells fail to induce Th17 polarization of naive T cells but do stimulate Th1 and Th17 memory responses. J. Immunol., 183(4):2242-51.
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