Transduction of cancer cell lines

The neuroblastoma cell line JF was kindly provided by Malcolm K. Brenner, Houston, USA. The Ewing’s sarcoma cell line TC-71 was obtained from DSMZ (Braunschweig, Germany). The prostate cancer cell line PC3 as well as the CHO cell line were purchased from American Type Culture Collection. For the bioluminescence based killing assay and the in vivo analysis JF and TC-71 cells were transduced to express the gene encoding firefly luciferase (JF Luc, TC-71 luc). Transduction was performed using a lentiviral packaging system as described previously [39 (link)]. PC3 cells expressing firefly luciferase were described previously [36 (link)]. All these cell lines were cultured in RPMI 1640 medium completed with 10% FCS, 100 U/ml penicillin and 100 µg/ml streptomycin, 2 mM N-acetyl-L-alanyl-L-glutamine, 1% non-essential amino acids and 1 mM sodium pyruvate (Biochrom). Human Embryonic Kidney cells HEK293T (ATCC CRL-11268) used for production of the lentiviral particles were cultured in DMEM medium supplemented with 10% FCS, 100 U/ml penicillin and 100 μg/ml streptomycin, 1% non-essential amino acids (Biochrom). Cells were maintained at 37°C in a humidified atmosphere of 5% CO₂.