Cell Culture and Viral Vectors for Cancer Research

The human lung fibroblast WI38 cell line, osteosarcoma cell line SAOS-2 (p53 null and truncated RB1), and the pancreatic cancer cell lines, BxPC-3, AsPC-1, MIA PaCa-2, and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA). WI38 cells were grown in Hyclone MEM/EBSS (ThermoFisher Scientific, Waltham, MA) media supplemented with 10% research grade fetal bovine serum (FBS) (PAA Laboratories, Dartmouth, MA) and 1% Penicillin Streptomycin (Corning, Corning, NY) and SAOS-2, MIA PaCa-2, and PANC-1 cells were grown in Hyclone High Glucose DMEM (ThermoFisher Scientific, Waltham, MA) supplemented with 10% FBS and 1% Penicillin Streptomycin. BxPC-3 and AsPC-1 were cultured in RPMI supplemented with 10% or 15% FBS (respectively) and 1% Penicillin Streptomycin. Cells were cultured at 37°C in a humidified 5% CO₂ incubator.
Ad.CMV (adenovirus with CMV promoter) and Ad.CMV.p53 (Adenovirus containing wild-type p53 gene under control of CMV promoter) viral vectors were generated using the AdEasy system (Carlsbad, CA). The Ad.CMV.pRb (Adenovirus containing RB1 gene cDNA under control of CMV promoter) vector was provided by Dr. Juan Fueyo (M.D. Anderson Cancer Center, The University of Texas). The Ad.GFP and Ad.GFP.RGS16 viruses were purchased from Vector Biolabs (Philadelphia, PA). Viruses were amplified and titered as previously described [81 (link)-83 (link)].