Expansion and Activation of T Cells

Primary T cells were cultured with IL-2 (50 ng/ml) and ImmunoCult Human CD3/CD28/CD2 T Cell Activator (25 μl/ml, 10970, STEMCELL Technologies) from PBMCs. T cells were expanded according to the manufacturer's protocol (58 (link), 81 ). In U-bottom 96 wells plate(3799, Corning), 1.2 × 10⁶ T cells were cocultured with 1.5 × 10⁵ BJAB cells (8:1) under different conditions. Total cells were treated with cell activation cocktail (with Brefeldin A) (423304, BioLegend) for 6 h before collected. After 72 h, cell cultures were stained for surface markers with CD3, CD4, and CD8a in PBS (with 2.5% FBS).

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Zhang X., Li Z., Peng Q., Liu C., Wu Y., Wen Y., Zheng R., Xu C., Tian J., Zheng X., Yan Q., Wang J, & Ma J. (2024). Epstein-Barr virus suppresses N6-methyladenosine modification of TLR9 to promote immune evasion. The Journal of Biological Chemistry, 300(5), 107226.

Publication 2024

ImmunoCult Human CD3/CD28/CD2 T Cell Activator U-bottom 96 wells plate cell activation cocktail (with Brefeldin A)

Corresponding Organization : Changzhi Medical College

Other organizations : Guilin Medical University, Xiangya Hospital Central South University

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Variable analysis

independent variables

Concentration of IL-2 (50 ng/ml)
Concentration of ImmunoCult Human CD3/CD28/CD2 T Cell Activator (25 μl/ml)

dependent variables

T cell expansion after 58 days
T cell expansion after 81 days
T cell activation (measured by cell surface markers CD3, CD4, and CD8a)

control variables

Culturing primary T cells from PBMCs
Culturing in U-bottom 96 wells plate
Ratio of T cells to BJAB cells (8:1)
Cell activation cocktail (with Brefeldin A) treatment for 6 hours before collection
Cell culture duration of 72 hours

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