Cell lines and cell culture

Tissue Factor (TF) expressing human breast cancer cell lines T47D, MDAMB231 and MCF7 were purchased from the American Type Culture Collection (ATCC) and verified as mycoplasma free. MCF7 cells stably transfected to overexpress TF and matched empty-vector control were provided Dr Henri Versteeg from Leiden University Medical Centre (co-author). Cell lines were maintained in adherent culture conditions at 37oC at atmospheric pressure in 5% (v/v) carbon dioxide/air. MCF7 and T47D cells lines were cultured in complete DMEM medium (DMEM/10%foetal calf serum/2 mM L-glutamine/PenStrep) and the MDAMB-231 cell line was cultured in complete RPMI-1640 medium (RPMI/10% FCS/1% Sodium pyruvate/2 mM L-glutamine/PenStrep).