T Cell Expansion and Characterization

T cells induced on DLL4 proteins were seeded into 48-well plates at a density of 1–10 × 10⁴ cells/well and stimulated with OKT3 anti-CD3 antibody (eBioscience, 500 ng/mL) in alpha-MEM supplemented with 15% FBS, 50 μg/mL PAA, 1% PSG, 10 ng/mL rhIL-7, and 10 nM dexamethasone (DEXART, Fuji Pharma) for 3 days (days 21–24).¹⁴ (link)^,36 (link) OKT3 and dexamethasone were removed by changing the medium (alpha-MEM supplemented with 15% FBS, 50 μg/mL PAA, 1% PSG, and 10 ng/mL rhIL-7), and cells were incubated for another 7 days (days 24–31). Anti-CD8β-PE-Cy7, anti-CD8α-APC, anti-CD45-APC-Cy7, anti-CD4-BV421, and anti-CD3-BV510 antibodies were used for FACS analysis.