Purified T cell Sample Clauses

Purified T cell. CFSE proliferation assay. Purified CD4 T cells were purified as described and resuspended in 107 cells/ml in PBS. CFSE was added at 5μM and incubated for 5-10 mins at 37⁰C. Cells were then washed in complete RPMI media and resuspended at 2x106 cells/ml in x-vivo. Cells were plated at 105/well in 96 well U-bottom plates in the relevant conditions and stimulated with 0.06μl of anti-CD3/28 dynobeads™ (Invitrogen Life Sciences). Following 6 days of proliferation cells were transferred to FACS tubes and assayed for CFSE dilution in the FITC channel of a FACS CantoII.
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