Caco 2

Human colorectal adenocarcinoma cells (CaCo-2), metastatic SW-620 colon cancer cells, hepatocellular carcinoma cell lines (Hep-3B, HLF and HLE cells), liver hepatoma cells (PLC-5), and gastric carcinoma (derived from metastatic site) N-87 cells were purchased from ATCC. Human gastric carcinoma (derived from metastatic site) cell line (HGC-27) was purchased from Sigma-Aldrich. Human intrahepatic cholangiocellular carcinoma cell line (HUCCT-1) was purchased from JCRB Cell Bank derived from ascitic fluid. All cell lines were cultivated in according to retailer protocols. Briefly, colon cancer cell lines (CaCo-2 and SW-620) were cultured in 10% of Inactivated Fetal Bovine Serum (FBS) exosomes free (Euroclone) in Dulbecco’s Modified Eagle’s Medium (DMEM) with sodium pyruvate, 4.5 g/L glucose (GIBCO), 4mM L-Glutamine (GIBCO) and 5 mL Pen-Strep (penicillin 10,000 u/mL, streptomycin 10,000 u/mL, Lonza Biowhittaker). Gastric (N-87 and HGC-27), hepatocellular carcinoma (Hep-3B, HLE and HLF), hepatoma (PLC-5), and intrahepatic cholangiocellular carcinoma (HUCCT-1) cell lines were cultured with Roswell Park Memorial Institute (RPMI) medium (GIBCO) supplemented with 10% of FBS exosomes free, sodium pyruvate, 4.5 g/L glucose, 4mM L-Glutamine, and 5 mL Pen-Strep.Cells were grown until reaching semi-confluence, in a humidified incubator at 37 °C with an atmosphere containing 5% of CO₂.

For this study we used two colon cancer cell lines with different degrees of differentiation, Caco-2 as a model for the differentiated phenotype and HCT116 for the cell type with a low degree of differentiation [34 (link)]. The lines were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA). HCT116 and Caco-2, were cultured at 37 °C in DMEM medium containing 10% and 20% Fetal Bovine Serum (FBS) respectively, 100 U/mL penicillin/streptomycin and 2 mM L-glutamine (EuroClone, Pero, MI, Italy). For the experiments, in cells grown at approximately 75% confluence, the cells were treated with dried extracts from 1 mL Olea europaea L. cvs. Leccino, Carboncella and Tortiglione olives, for 72 hours’ exposure time at different dilutions (1:100; 1:500; 1:1000; 1:10,000).