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StemXVivo Serum-Free Dendritic Cell Base Media

R&D Systems, part of Bio-Techne | Catalog # CCM003

R&D Systems, part of Bio-Techne

Key Product Details

Differentiation of CD14+ Monocytes into Dendritic Cells. 
(3)

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
    View Graphic Procedure
 

 

Reagents Provided

Reagents supplied in the StemXVivo® Serum-Free Dendritic Cell Base Media (Catalog # CCM003):

  • 250 mL StemXVivo® Serum-Free Dendritic Cell Base Media

 

Other Supplies Required

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

 

Procedure Overview

Differentiation of CD14+ Monocytes into Monocyte-derived Dendritic Cells Using StemXVivo® Serum-Free Dendritic Cell Base Media (Catalog # CCM003)

  1. Thaw Human StemXVivo® Serum-Free Dendritic Cell Base Media.
  2. Mix the thawed media by vortexing and warm the media in a 37 °C incubator.
  3. Prepare the CD14+ cells from peripheral blood mononuclear cells.
  4. Resuspend the purified CD14+ cells in Human StemXVivo® Serum-Free Dendritic Cell Base Media.
  5. Perform a cell count and adjust the cell density to 1 x 106 cells/mL.
  6. Supplement the media with Gentamicin, GM-CSF, and IL-4.
  7. Add the cell suspension to the tissue culture flask or tissue culture plate.
  8. Incubate the cells in a 37 °C and 5% CO2 humidified incubator for 7 days.
  9. Change the media every three days.
  10. Observe immature monocyte-derived dendritic cells (MoDCs) from day 5 to day 7.
  11. Induce maturation of MoDCs by adding LPS to the cell suspension on day 7 and incubating for 48 hours.
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Product Documents for StemXVivo Serum-Free Dendritic Cell Base Media

Certificate of Analysis

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