Neutralized n acetyl l cysteine

Primary CD4+ and CD8+ T cells were isolated from anonymous donor blood after apheresis by negative selection (STEMCELL Technologies #15062 & 15063). Blood was obtained from Blood Centers of the Pacific (San Francisco, CA) as approved by the University Institutional Review Board. T cells were cryopreserved in RPMI-1640 (UCSF cell culture core) with 20% human AB serum (Valley Biomedical Inc., #HP1022) and 10% DMSO. After thawing, T cells were cultured in human T cell medium consisting of X-VIVO 15 (Lonza #04-418Q), 5% Human AB serum and 10 mM neutralized N-acetyl L-Cysteine (Sigma-Aldrich #A9165) supplemented with 30 units/mL IL-2 (NCI BRB Preclinical Repository) for all experiments.

Primary CD4⁺ and CD8⁺ T cells were isolated from donor blood after apheresis by negative selection (STEMCELL Technologies, nos. 15062 and 15063). Blood was obtained from Blood Centers of the Pacific, as approved by the University of California San Francisco (UCSF) Institutional Review Board. T cells were cryopreserved in RPMI 1640 (UCSF cell culture core) with 20% human AB serum (Valley Biomedical, no. HP1022) and 10% dimethyl sulfoxide. After thawing, T cells were cultured in human T cell medium consisting of X-VIVO 15 (Lonza, no. 04–418Q), 5% human AB serum, and 10 mM neutralized N-acetyl L-cysteine (Sigma-Aldrich, no. A9165) supplemented with IL-2 (30 U/ml; National Cancer Institute Biological Resources Branch Preclinical Repository) for all experiments except for the IncuCyte experiments. IncuCyte experiments were cultured in RPMI 1640 with 5% human AB serum supplemented with IL-2 (30 U/ml).