2.3. Human CAR-T cells generation
Human peripheral blood mononuclear
cells (PBMCs) samples were
obtained from healthy donors according to the manufacturer’s protocol.
Written informed consents were obtained from all donors. PBMC were
stimulated with anti-CD3/CD28 for 48 h. Then, the activated T cells were
infected with the CAR-encoding lentiviral supernatant. 24h after
lentiviral infection, the lentiviral supernatant was replaced with fresh
medium containing RPMI-1640 and supplemented with 10% FBS (Gibco),
200U/ml IL-2, 25 mM HEPPES, 55 μM 2-M, 100 U/ml penicillin and 100 μg/ml
streptomycin. The infected T cells were cultured for 7 days.