For growth curve, cell viability was monitored by the IncuCyte live cell analysis system (Essen BioScience, Ann Arbor, MI, USA). To assess the cell proliferation, CRC cells were equally seeded into fresh 12-well plates at a density of 1 × 105 (link) cells. Cells were treated with different indicated concentrations of C.B CM. After placing the cell in a cell incubator, the IncuCyte live cell analysis system analyze the photographs at 9 random fields to obtain growth curve.
For IC50 calculation, cell viability was determined by Cell Counting Kit-8 (K1080, Apexbio Technology LLC, Houston, TX, USA). CRC cells were seeded into 96-well plates for 24 h. The cells were treated with indicated doses of C.B CM, 5-FU, and the combination of C.B CM and 5-FU for 48–72 h. Following that, 10% (v/v) CCK-8 solution was prepared and added to every well, and cells were plated into a cell incubator for 24 h. Optical density was measured at 450 nm using Epoch™ Multi-Volume Spectrophotometer and Take3™ (BioTek, Winooski, VT, USA). Cell viability was normalized as a percentage of the negative controls treated with culture medium. IC50 values were calculated by using linear regression equations obtained from the concentration vs percentage of viable cells.
For IC50 calculation, cell viability was determined by Cell Counting Kit-8 (K1080, Apexbio Technology LLC, Houston, TX, USA). CRC cells were seeded into 96-well plates for 24 h. The cells were treated with indicated doses of C.B CM, 5-FU, and the combination of C.B CM and 5-FU for 48–72 h. Following that, 10% (v/v) CCK-8 solution was prepared and added to every well, and cells were plated into a cell incubator for 24 h. Optical density was measured at 450 nm using Epoch™ Multi-Volume Spectrophotometer and Take3™ (BioTek, Winooski, VT, USA). Cell viability was normalized as a percentage of the negative controls treated with culture medium. IC50 values were calculated by using linear regression equations obtained from the concentration vs percentage of viable cells.
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