Hct 116

A375-C5 (melanoma) and MCF-7 (breast adenocarcinoma) cell lines were obtained from the European Collection of Cell Culture, UK, and were grown in RPMI-1640 medium (Roswell Park Memorial Institute, Biochrom, Cambridge, UK) with 5% inactivated FBS (fetal bovine serum, Biochrom). HCT-116 (colon adenocarcinoma) cells were grown in DMEM medium (Dulbecco’s Modified Eagle’s, Biochrom) with 5% inactivated FBS. The non-tumor human pulmonary alveolar epithelial cells (HPAEpiC) were obtained from ScienCell Research Laboratories and grown in DMEM medium supplemented with 10% FBS and 1% non-essential amino acids (Sigma-Aldrich Co., Saint Louis, MO, USA). All cell lines were maintained in a humidified incubator (Hera Cell, Heraeus, Hanau, Germany) at 37 °C and with 5% CO₂, and cell viability was regularly monitored using the exclusion trypan blue dye.

All cell lines, with exception of HUVECs, were from the American Type Culture Collection (ATCC, Manassas, VA, USA). HCT-116 and HT-29 were kept at 37 °C, humidified 5% CO₂ incubators and cultured with McCoys medium supplemented with 10% fetal bovine serum (FBS) (Biochrom, Berlin, Germany) and 1% Pen/Strep (Gibco, Thermo Fischer Scientific, Waltham, MA, USA). SW-1463 cells were kept at 37 °C, humidified atmospheric-air incubators and cultured with Leibovitz’s L-15 medium supplemented with 10% FBS (Biochrom) and 1% Pen/Strep (Gibco). Primary, single-donor early passage HUVECs were kindly provided by Dr. Joanna Kalucka (Department of Biomedicine, Aarhus University, Aarhus, Denmark) and cultured at 37 °C, 5% CO₂ humidified incubators with Endopan 300SL medium (PAN Biotech, Aidenbach, Germany). Cells between passages five to twelve (HCT-116, HT-29, SW-1463) and passages two to three (HUVECs) were used for experiments. Normal colon epithelial cells (passages 1 and 2) were cultures with DMEM/F12 medium (ATCC) supplemented with 10% non-inactivated FBS (Biochrom), 10 ng/mL cholera toxin (Sigma), 0.005 mg/mL insulin (Gibco), 0.005 mg/mL transferrin (Gibco), 100 ng/mL hydrocortisone (Sigma) and 20 ng/mL human EGF (Gibco). The final DMSO concentration experienced by cells in vitro was ≤ 0.25%.