Culturing Human Cancer Cell Lines

Human oral cancer cell line OC3-IV2 cells have been described [9 (link),25 (link),26 (link)] and were cultured in a 1:1 ratio of Dulbecco’s modified Eagle medium (DMEM; Invitrogen) and keratinocyte serum-free medium (KSFM; Invitrogen) containing 10% (v/v) fetal bovine serum (Invitrogen), 1% (v/v) l-glutamine (Invitrogen), and 1% (v/v) antibiotic-antimycotic (Invitrogen). Human primary glioblastoma cell line U87 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and grown in DMEM medium supplemented with 10% (v/v) fetal bovine serum, 1% (v/v) Minimum Essential Medium (MEM) non-essential amino acids solution (Invitrogen), and 1% (v/v) penicillin/streptomycin (Invitrogen). Human prostate cancer cell line PC3 cells, human cervical cancer cell line HeLa cells, and 293T cells were obtained from the ATCC and maintained in DMEM medium supplemented with 10% (v/v) fetal bovine serum, 1% (v/v) l-glutamine, and 1% (v/v) antibiotic-antimycotic. Human non-small cell lung carcinoma cell line H1299 cells were obtained from the ATCC and were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Invitrogen) containing 10% (v/v) fetal bovine serum, 1% (v/v) l-glutamine, and 1% (v/v) antibiotic-antimycotic. Cells were maintained in a humidified atmosphere containing 5% CO₂ at 37 °C, and the culture medium was replaced every 2 days.