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StemXVivo Serum-Free Dendritic Cell Base Media

Key Product Details

Differentiation of CD14+ Monocytes into Dendritic Cells. 
(3)
Phenotypic Analysis of Cultured Monocyte-derived Dendritic Cells Before and After LPS-induced Maturation. 
Mature Monocyte-derived Dendritic Cells Induce Proliferation of Allogenic T Cells.
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CCM003

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 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 (Tocris Catalog # 4131)

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)

Thaw Human StemXVivo® Serum-Free Dendritic Cell Base Media.

Mix the thawed media by vortexing and warm the media in a 37 °C incubator.

Human StemXVivo Serum-Free Dendritic Cell Base Media

Prepare the CD14+ cells from peripheral blood mononuclear cells.

Resuspend the purified CD14+ cells in Human StemXVivo® Serum-Free Dendritic Cell Base Media.

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.

Perform a cell count

Incubate the cells in a 37 °C and 5% CO2 humidified incubator for 7 days.

Change the media every three days.

Incubate the cells

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.

Add LPS to the cell suspension

Citations (2)

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    StemXVivo Serum-Free Dendritic Cell Base Media Serum CCM003

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