Melanoma Cell Line Culture Protocol

PLX4032 (also known as RG7204 or RO5185426) was obtained under a materials transfer agreement (MTA) with Plexxikon (Berkeley, CA) and dissolved in DMSO (Fisher Scientific, Morristown, NJ) to a stock concentration of 10 mM. SKMEL28 was obtained from American Type Culture Collection (ATCC, Rockville, MD), and the remaining human melanoma cell lines (M series) were established from patient's biopsies under UCLA IRB approval #02-08-067. Cells were cultured in RPMI 1640 with L-glutamine (Mediatech Inc., Manassas, VA) containing 10% (unless noted, all percentages represent volume to volume) fetal bovine serum (FBS, Omega Scientific, Tarzana, CA) and 1% penicillin, streptomycin, and amphotericin (Omega Scientific). All cell lines were mycoplasma free when periodically tested using a Mycoalert assay (Lonza, Rockland, ME).