Mem neaa

Six human squamous cell carcicnoma (SCC) cell lines (UM-SCC-74A, UM-SCC-1, UM-SCC-11A, UM-SCC-47, UD-SCC-2, and ATCC CAL 27) were used and cultured in Dulbecco modified Eagle medium (DMEM, Gibco/Invitrogen, Carlsbad, CA) supplemented with 10% heat inactivated fetal bovine serum (FBS, Gibco/Invitrogen), 50 μg/ml of penicillin G, 50 μg/ml of streptomycin sulfate, and 1× MEM NEAA (Gibco/ Invitrogen). Vero (African green monkey kidney cells, ATCC) cells were maintained in DMEM containing 10% FBS without 1× MEM NEAA. All cell lines were maintained at 37 °C in a humidified atmosphere at 5% CO₂.
The construction and generation of both control rHSVQ1 and RAMBO (Rapid Antiangiogenesis Mediated By Oncolytic Virus) viruses have been previously described.^{30 (link)} Viruses were propagated in Vero cells, purified, and infectious virus titers (plaque forming unit per ml (pfu/ml)) were determined by standard plaque forming unit assay on Vero cells.^{31 (link),32 (link)}

Jurkat NFAT-luc acute lymphatic leukemia cell line, expressing a luciferase reporter under the control of an NFAT-dependent promoter was purchased from Promega Corporation. Cells were cultured in RPMI medium with HEPES, GlutaMAX, MEM NEAA, Sodium pyruvate, 10% fetal calf serum (FCS) and 200 μg/mL hygromycin B. The target cell line was purchased from ATCC and cultured in IMDM with HEPES, l-glutamine and 10% FCS. A549 lung carcinoma cell line was generated in-house by lentiviral transduction with human full length TAA2. Cells were cultured in DMEM with high glucose, l-glutamine, pyruvate, 10% FCS and 4 μg/mL Puromycin dihydrochloride. All cells were passaged twice a week.