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StemXVivo Serum-Free Human T Cell Base Media

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Proliferative Response of Cultured CD3/CD28 Primed T cells in Human StemXVivo® Serum-Free T Cell Base Media. 
(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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CCM010
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Assay Procedure

Refer to the product datasheet for complete product details.

CD3/CD28-primed T lymphocytes can be cultured ex vivo in StemXVivo® Serum-Free T Cell Base Media using the following procedure:

  • Coat a 96-well microplate with CD3 and CD28 antibodies
  • Resuspend CD3+ T cells in StemXVivo® Serum-Free T Cell Base Media
  • Add CD3+ T cells to the microplate
  • Assess proliferation after 3-5 days
 

 

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Reagents Provided

Reagent Supplied in StemXVivo® Serum-Free T Cell Base Media (Catalog # CCM010):

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

 

Other Supplies Required

Reagents

  • CD3+ lymphocytes isolated from PBMCs using MagCellect Human CD3+ T Cell Isolation Kit (Catalog # MAGH101), or Cell Enrichment Column (Catalog # HTCC)
  • Human CD3 monoclonal antibody (Catalog # MAB100)
  • Human CD28 polyclonal antibody (Catalog # AF-342-PB)
  • Penicillin-Streptomycin
  • Recombinant Human IL-2 (Catalog # 202-IL, optional)
  • [3H]-thymidine

Materials

  • 96 well v- or round-bottom microplate
  • 15 mL centrifuge tubes
  • Serological pipettes
  • Pipettes and pipette tips
  • Hemocytometer
  • Cell harvester

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge
  • Vortex Mixer
  • Inverted microscope
  • Beta-scintillation counter

 

Procedure Overview

Protocol for Culturing CD3/CD28-Primed T Lymphocytes Ex Vivo using StemXVivo® Serum-free T Cell Base Media (Catalog # CCM010)

Preparation of the CD3/CD28 coated 96-well microplate

Add working solution of CD3 and CD28 antibody to each well of a 96-well microplate.

Incubate for 90 minutes at 37 °C or at 2 °C to 8 °C overnight.

Wash the wells with PBS three times

Preparation of the CD3/CD28 coated 96-well microplate

Thawing StemXVivo® Serum-Free T Cell Base Media and Plating CD3+ T Cells

Thaw the Human StemXVivo® Serum-Free T Cell Base Media

Prepare CD3+ T cells from peripheral blood mononuclear cells.

Thawing StemXVivo Serum-Free T Cell Base Media and Plating CD3+ T Cells

Resuspend the purified CD3+ T cells in Human StemXVivo® Serum-Free T Cell Base Media.

Resuspend the purified CD3+

Perform a cell count and adjust the cell density to 1 x 106 cells/mL.

Perform a cell count

Add the CD3+ T cells to each well at 2 x 105 cells in 0.2 mL.

Place the plate in a humidified 37° C and 5% CO2 incubator for 3-5 days for short-term stimulation.

Add the CD3+ T cells to each well

Assess the proliferation of CD3/CD28-primed T cells by adding [3H]-thymidine to each well during the last 18 hours of a 3-5 day period.

Harvest cells using a cell harvester and measure the cpm (counts per minute) with a beta-scintillation counter.

Assess the proliferation of CD3/CD28-primed T cells

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Product Documents for StemXVivo Serum-Free Human T Cell Base Media

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