This application protocol describes use of the Treg Expansion Kit, human, which is designed to efficiently expand Treg cells that maintain FoxP3 expression after isolation with the CD4+CD25+ Regulatory T Cell Isolation Kit, human. It includes a complete protocol for your Treg cell workflow, describing human Treg cell isolation from peripheral blood mononuclear cells (PBMCs), their in vitro expansion, and subsequent flow cytometry analysis.
1× PBS (pH 7.2)
+ 0.5 % BSA
+ 2 mM EDTA
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Example:
For 1 sample with 1×106 Treg cells: prepare 1 mL of fixation/permeabilization solution and 4 mL of 1× Permeabilization Buffer.
The expansion medium can be stored for up to 10 days under sterile conditions at 4 °C.
500 mL TexMACS™ Medium
+ 500 IU/mL Human IL-2 IS, premium grade
+ 5 % human AB serum
+ 0.01 mM β-ME
+ 1 % Penicillin/streptomycin
+ (Optional) 100 nmol/L rapamycin
▲ Note: Rapamycin has been shown to increase the frequency of FoxP3+ Treg cells during expansion (Battaglia, M. et al. (2005) Blood 105: 4743–4748).
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