Fresh blood was collected from healthy volunteers after obtaining informed consent from the review committee of China Pharmaceutical University. Peripheral blood mononuclear cells were isolated from fresh blood by gradient centrifugation with use of Lymphoprep™ (Axis‐Shield, Norseland). T cells were selectively enriched with CD3+ separation beads (Miltenyi Biotec Inc, Auburn, CA, USA). Isolated T cells were cultured in X‐VIVO15 culture medium (Lonza, Switzerland) supplemented with 5% human AB serum (Valley Biomedical Inc, Winchester, VA, USA), 10 mM N‐acetyl l ‐cysteine (Sigma Aldrich, St. Louis, MO, USA), and 300 IU/mL recombinant human IL‐2 (PeproTech, Rocky Hill, CT, USA).
Human pancreatic cancer cell AsPC‐1 and BxPC‐3, colorectal cancer cell HT‐29 and gastric cancer cell MGC803 were obtained from American Type Culture Collection. AsPC‐1 cells were cultured in RPMI1640 culture (Hyclone, Logan, UT, USA) and PANC‐1, HT‐29, and MGC‐803 cells were cultured in DMEM culture (Hyclone). All the tumor cultures were supplemented with 10% fetal bovine serum (FBS) (Gibco, Gaithersburg, MD, USA), 2 mmol/L Glutamine (Gibco), 100 U/mL penicillin, and 100 µg/mL streptomycin (Sangong Biotech, Shanghai, China).
Human pancreatic cancer cell AsPC‐1 and BxPC‐3, colorectal cancer cell HT‐29 and gastric cancer cell MGC803 were obtained from American Type Culture Collection. AsPC‐1 cells were cultured in RPMI1640 culture (Hyclone, Logan, UT, USA) and PANC‐1, HT‐29, and MGC‐803 cells were cultured in DMEM culture (Hyclone). All the tumor cultures were supplemented with 10% fetal bovine serum (FBS) (Gibco, Gaithersburg, MD, USA), 2 mmol/L Glutamine (Gibco), 100 U/mL penicillin, and 100 µg/mL streptomycin (Sangong Biotech, Shanghai, China).