Ham f10 medium

The cell lines were derived from melanoma metastasis by the Laboratory of Clinical and Experimental Oncology at the Jules Bordet Institute (Brussels, Belgium). We worked with three cell lines harbouring different mutations and sensitive to specific targeted therapy (MM074 (V600EBRAF), MM161 (Q61RNRAS) and HBL (D820YcKIT)) and their corresponding cells with acquired resistance to the drugs (-R) developed by chronic treatment with increasing concentrations of the inhibitors. Cells were grown in Ham-F10 medium (Lonza, Bâle, Switzerland) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both from Life Technologie, Carlsbad, CA, USA). Cells were confirmed to be free of mycoplasma contamination by qPCR analysis.

Three cell lines harboring different mutations were selected: HBL (^D820YcKIT), MM074 (^V600EBRAF) and MM161 (^Q61RNRAS). These lines are sensitive to targeted therapies, but we also worked with their resistant counterparts. To develop these resistances, cells were chronically exposed to increasing doses of targeted therapies for 12 weeks (0.01 µM during Week 1 and 2; 0.05 µM during Week 3 and 4; 0.1 µM during Week 5 and 6; 0.5 µM during Week 7 and 8; 1 µM during Week 9 and 10; 2 µM during Week 11 and 12). This model mimics what happens in patients who develop resistances to targeted therapies. Both sensitive and resistant cell lines were provided by the Laboratory of Oncology and Experimental Surgery at Institut J. Bordet, Université libre de Bruxelles (Brussels, Belgium).
Cells were grown in Ham-F10 medium (Lonza, Bâle, Switzerland) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both from Life Technologies, Carlsbad, CA, USA) at 37 °C in a humidified 95% air and 5% CO₂ atmosphere. The cultures were confirmed to be free of mycoplasma contamination using PCR-based detection.

Metastatic melanoma cell lines were derived from tumors by the Laboratory of Oncology and Experimental Surgery at Institut J. Bordet, Université libre de Bruxelles (Brussels, Belgium). For the work, we selected 3 cell lines harboring different mutations: HBL (^D820YcKIT), MM074 (^V600EBRAF), and MM161 (^Q61RNRAS). These lines are sensitive to targeted therapies, but we also worked with their resistant counterparts. To develop these resistances, cells have been chronically exposed to increasing doses of targeted therapies for 12 weeks (0.01–1 µM) by M. Krayem and A. Najem. This model mimics what happened in patients developing resistances to targeted therapies. Acute drug exposure effects were generated by exposing sensitive cells to 1 µM of their corresponding drug during 72 h. Due to Dabrafenib’s impact on the MM074-sensitive cell density, a shortened time-point of 24 h of exposure was also considered.
Cells were grown in Ham-F10 medium (Lonza, Bâle, Switzerland) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both from Life Technologie, Carlsbad, CA, USA) at 37 °C in a humidified 95% air and 5% CO₂ atmosphere. The cultures were determined to be free of Mycoplasma contamination using PCR-based detection.