Different groups of SW480 and HCT116 cells were treated with either BSA or 400 μM oleic acid (OA) for 6 hours, then washed and incubated with phenol red‐free DMEM complexed with 1% BSA for additional 24 hours. The supernatant was collected and subjected to subsequent analyses. FFA from cell culture medium was measured using the EnzyChromTM Free Fatty Acid Assay Kit, according to the manufacturer's instructions. FFA was calculated from a standard curve for each assay, and the data were normalized to total protein.