CellXVivo Human Treg Cell Differentiation Kit
R&D Systems, part of Bio-Techne | Catalog # CDK006
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, mature human CD4+ T cells can be differentiated into Tregs using the following procedure:
- Coat a plate with Mouse Anti-Human CD3 Antibody
- Isolate naïve CD4+ T cells from Human PBMCs
- Culture naïve CD4+ T cells in Human Treg Differentiation Media for 5 days
- Verify differentiation into Treg cells by flow cytometry
Reagents Supplied in the CellXVivo™ Human T (Treg) Cell Differentiation Kit (Catalog # CDK006):
- Mouse Anti-Human CD3 Antibody
- Human Treg Reagent 1
- Human Treg Reagent 2
- Human Treg Reagent 3
- Human Treg Reagent 4
- Reconstitution Buffer 1
- Reconstitution Buffer 2
- Wash Buffer (20X)
Reagents
- Ficoll-Hypaque™
- MagCellect™ Human Naïve CD4+ T Cell Isolation Kit (R&D Systems, Catalog # MAGH115, or equivalent)
- X-VIVOTM15 Chemically Defined, Serum-free Hematopoietic Cell Medium (Lonza, or equivalent)
Equipment
- Tissue culture flasks and/or plates
- Sterile deionized water
- Microscope
- Hemocytometer
- 37 °C and 5% CO2 incubator
- Centrifuge
- Pipettes and pipette tips
Coat wells of a 24-well plate with Mouse Anti-Human CD3 Antibody.
Isolate PBMCs from human blood.
Isolate human naïve CD4+ T cells from PBMCs (e.g., using magnetic cell selection).
Perform a cell count.
Suspend 1 - 2 x 105 naïve CD4+ T cells/mL in Human Treg Differentiation Media.
Culture the cells on plates pre-coated with CD3 antibody for 5 days.
Verify Treg cell differentiation by analyzing marker expression using flow cytometry.
Mature Treg cells are now ready for further downstream applications.