PBMC-derived T cells were activated with 1 µg/mL of Ultra-LEAF™ anti-human CD3 antibody (Cat. No. 317347), 1 µg/mL of Ultra-LEAF™ anti-human CD28 antibody (Cat. No. 302943) in IMDM basal media supplemented with 200 IU/mL Recombinant Human IL-2 (Cat. No. 791908) and media additives as indicated. On day 10, flow cytometry was used to determine specific T cell populations by making use of anti-human CD62L and anti-human CD45RO antibodies.
PBMC-derived T cells were activated with 1 µg/mL of Ultra-LEAF™ anti-human CD3 antibody (Cat. No. 317347), 1 µg/mL of Ultra-LEAF™ anti-human CD28 antibody (Cat. No. 302943) in IMDM basal media supplemented with 200 IU/mL Recombinant Human IL-2 (Cat. No. 791908) and media additives as indicated. (A) Cell expansion was determined on day 4, 6, 8, and end of culture (day 11). (B) Flow cytometry was used to determine specific T cell populations, on day 11, by anti-human CD4 (Cat. No. 317408) and anti-human CD8 (Cat. No. 344722) staining.
Our phenotypic analysis showed that the cells expanded in either the xeno-free or CD serum substitutes exhibited different functional properties. Using the same protocol, the xeno-free serum substitute tends to expand a greater proportion of naïve CD3+ T cell and regulatory T cell (Treg) populations, while the CD formulation preferentially expands central memory T cells and naïve T cells.
Human naïve CD4+ T cells were stimulated with Ultra-LEAF™ anti-human CD3 and Ultra-LEAF™ anti-human CD28 antibodies (1 µg/mL), and Recombinant Human IL-2 (200 IU/mL) alone (data not shown) or with the additions of Recombinant Human TGF-β (5 ng/mL) for 6 days in the presence of human AB serum, Cell-Vive™ T Cell CD Serum Substitute, GMP, or Cell-ViveTM T-NK Xeno-Free Serum Substitute. ANOVA was used to determine the effect of serum additives on Treg levels.*p<0.05
Researchers can now select the serum substitute formulation that works best for their application and desired cell type. For any application inquiries you may have, please contact BioLegend Technical Support. Our team is ready to help.
Follow Us