Chemosensitivity

For viability evaluation after chemotherapeutic treatment 5000 cells were seeded into 96-well plates in sextuplicates. One day post seeding cells were treated with following chemotherapeutics 5-Fluorouracil, Etoposide, Gemcitabine, Oxaliplatin, Irinotecan and Methotrexate. Clinically widely used Ifosfamide was also evaluated but it proved to be unsuitable for in vitro experiments as it needs to be metabolized within the liver to produce the active metabolites. Untreated MaS-3 cells were used as a negative control. Viability of the cells was assed 48 and 72 h post treatment via MTT assay. Briefly, 20 µl of 5 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) solution was added to each well and incubated for 4 h at 37 °C. The MTT formazan was dissolved in DMSO and absorbance was measured at 560 nm with reference at 670 nm and expressed as relative values compared to untreated control cells (Tecan infinite M200, Tecan Group).