GMP Cloudz Human T Cell Activation KitGMP

Expansion of Human T Cells using GMP Cloudz™ Human T Cell Activation Kit Primary human CD3⁺ cells were activated with GMP Cloudz™ Human T-Cell Activation Kit and cultured for 9 days in ExCellerate™ T Cell Expansion Media (R&D Systems, Catalog # CCM030) and 20 ng/mL IL-2 (R&D systems, Catalog # 202-GMP). A. Cell counts were performed to determine fold expansion compared to the Day 0 seeding density (0.25 x 10⁶ cells/mL).

Viability of Human T Cells using Cloudz™ Human T Cell Activation Kit. Primary human CD3⁺ cells were activated with GMP Cloudz™ Human T-Cell Activation Kit and cultured for 9 days in ExCellerate™ T Cell Expansion Media (R&D Systems, Catalog # CCM030) and 20 ng/mL IL-2 (R&D systems, Catalog # 202-GMP). Cell counts were performed to determine percent viability on days 5, 7, and 9 of the experiment compared to the Day 0 seeding density (0.25 x 10⁶ cells/mL).

Phenotypic analysis of CD25/CD69 Markers Primary human CD3⁺ cells were A. activated with GMP Cloudz™ Human T Cell Activation Kit and cultured for 48 hours in ExCellerate™ T Cell Expansion Media (R&D Systems, Catalog # CCM030) and 20 ng/mL IL-2 (R&D systems, Catalog #202-GMP). B. The negative control was performed without adding Cloudz T Cell microspheres. Cells were fluorescently stained with using CD25-PE (R&D Systems, Catalog # FAB1020P) and CD69-a488 (R&D Systems, Catalog # FAB23591G) and analyzed via flow cytometry. Dead cells and debris were excluded from the analysis on scatter signals and viability dye. Cells that were treated with GMP Cloudz™ Human T Cell Activation Kit showed an activation signal 48 hours after reagent addition.