*2.4. Cell Culture*

The CaCo-2 cell line is originally derived from a primary colon adenocarcinoma (Cancer Coli-2) and was established by Jorgen Fogh at the Sloan Kettering Cancer Research Institute, from a Caucasian male (ATCC). The CoLo-205 cell line has been established from ascites fluid obtained from a male patient with metastatic adenocarcinoma of the colon (ATCC). CaCo-2 cells were cultured in Eagle's Minimum Essential Medium, (GibCo), which was added with 10% of fetal bovine serum, 1% of penicillin/streptomycin and 1% of glutamine. For the CoLo-205 cell line, ATCC-formulated Roswell Park Memorial Institute (RPMI)-1640 Medium was employed, with the addition of 10% of fetal bovine serum (10%), 1% of penicillin/streptomycin (1%) and 1% of glutamine (1%). When the cell lines confluence was about 70%, the cell layer was rinsed with PBS, and trypsinized, for the subsequent in vitro experiments.
