Human acute T cell leukemia cell line Jurkat (American Type Culture Collection (ATCC), TIB-152), lung cancer cell line NCI-H292 (ATCC, CRL-1848) and ovarian cancer cell line SKOV3 (ATCC, HTB-77) was cultured in RPMI 1640 (Corning) supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin-streptomycin (Pen/Strep, Corning). Embryonic kidney cell line HEK293T (ATCC, CRL-3216), lung cancer cell line A549 (ATCC, CCL-185) and epithelioid cervix carcinoma cell line Hela (ATCC, CBP-60232) were cultured in Dulbecco’ s modified Eagle’s medium (Corning) supplemented with 10% FBS and 1% Pen/Strep. Tumor cells carrying the report gene firefly luciferase (SKOV3-Luc, NCI-H292-Luc and A549-Luc cells) were engineered as previously described.36 (link) The cells were maintained in a humidified atmosphere containing 5% CO2 at 37°C. The hypoxia treatment (1% O2, 5% CO2 and 94 % N2) was carried out in Smartor 118 mobile three-gas control system (China Innovation Instrument) or by adding different concentration of CoCl2 into the culture medium to simulated chemical hypoxia.
The recombinant type 5 adenovirus expressing tumor antigen Her2 with reporter firefly luciferase (Ad5-Her2) was contracted for production to Shanghai Genechem Co., Ltd.