Culturing MPNST Cell Lines for Drug Testing

Mouse-derived MPNST cell lines described previously [34 (link)] were cultured at 37 °C in a humidified atmosphere in 5% CO₂ in low pH DMEM (Thermo Fisher Scientific) supplemented with 10% FBS (Corning, lot #35070165) and 1% Penicillin Streptomycin (Thermo Fisher Scientific), unless otherwise indicated. Cell lines were verified to be free of Mycoplasma contamination every 6 months by PCR (ATCC). Human-derived MPNST cell lines were cultured as described previously [85 (link)]. Capmatinib (Novartis), trametinib (Novartis), everolimus (Selleckchem), and afuresertib (Selleckchem) solutions were prepared in DMSO. For human cell line [86 (link)] IC₅₀ experiments, drugs were purchased as 10 mM stock solutions (Selleckchem) and handled as previously described [85 (link)]. All cell culture experiments were performed 3 independent times, unless otherwise noted.