Hct15

DNA from the following 20 colon cancer cell lines was used to investigate promoter methylation status; Co115, Colo205, Colo320, EB, FRI, HCT15, HCT116, HT29, IS1, IS3, KM12, LS1034, LS174T, NCI-H508, RKO, SW480, SW620, SW948, TC71, and V9P. HCT15, HCT116, NCI-H508, RKO, SW620, and SW948 have been purchased from the American Type Culture Collection (ATCC), and Colo205 and KM12 from the Charles River Laboratories. The rest of the cell lines have kindly been provided by Professor Richard Hamelin (INSERM, Paris, France). DNA was isolated using either a standard phenol/chloroform protocol, or a magnetic beads approach (Maxwell^® 16 System, Promega). For each cell line, the same DNA stock was used for all analyses. All cell lines have been tested by short tandem repeat (STR) profiling using the AmpFLSTR Identifiler PCR Amplification Kit (Life Technologies). Commercial lines have been authenticated based on comparisons with available STR profiles from the American Type Culture Collection (ATCC) and the German Collection of Microorganisms and Cell cultures13 (link). All cell lines tested negative for mycoplasma infection (MycoAlert Mycoplasma Detection Assay and Lucetta Luminometer, Lonza).

Human colorectal cancer cell lines HT-29, HCT-15, and HCT-116 are cell lines from the U.S. National Cancer Institute’s NCI-60 tumor cell panel and were purchased from Charles River Laboratories (Charles River Laboratories Inc., New York, NY, USA). Normal human cell lines CCD-18 Co (non-malignant fibroblastic colon cells) and CCD-841 CoN (non-malignant epithelial colon cells) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). African green monkey kidney (Vero) cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany).
Tumor cells and Vero cells were maintained in high-glucose (4.5 g glucose per liter) Dulbecco’s Modified Eagle’s Medium (DMEM, D6429, Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal calf serum (FCS; Biochrom, Berlin, Germany). Normal human cell lines were grown in Alpha Modified Eagle’s Medium (Alpha-MEM, BE12-169F, Lonza, Verviers, Belgium) enriched with 10% FCS and 10 mM L-glutamine (Gibco, Paisley, Scotland, UK). Cell culture flasks were stored in humidified incubators at 37 °C under 5% CO₂.