Cv crystal violet based cell viability assays

The investigated cell lines, PC-3 (human prostate adenocarcinoma) and HT-29 (human colorectal adenocarcinoma) were purchased from ATCC (Manassas, VA, USA). The cell culture medium RPMI 1640, the supplements FCS and l-glutamine, as well as PBS and trypsin/EDTA were purchased from Capricorn Scientific GmbH (Ebsdorfergrund, Germany). Culture flasks, multi-well plates, and further cell culture plastics were purchased from Greiner Bio-One GmbH (Frickenhausen, Germany) and TPP (Trasadingen, Switzerland), respectively. Anti-proliferative and cytotoxic effects, respectively, of the compounds, were investigated by performing colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and CV (crystal violet)-based cell viability assays (Sigma-Aldrich, Taufkirchen, Germany), respectively.

Briefly, for the cytotoxicity assay, the human prostate cancer cell line PC-3 and the colon adenocarcinoma cancer cell line HT-29 (both from ATCC, Manassas, VA, USA) were used. The cell handling and assay techniques were in accordance with the method described by Khan et al. [34 (link)]. The extract was tested at the concentrations of 0.05 and 50 μg/mL. Anti-proliferative and cytotoxic effects of the extract were investigated by performing colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and CV (crystal violet)-based cell viability assays (Sigma-Aldrich, Taufkirchen, Germany) after 48 h treatment time, respectively. The absorbance was measured with an automated microplate reader at 540 nm with a reference wavelength of 670 nm. Digitonin (125 μM) was used as positive control, which was set for data normalization to 0% cell viability. The results are presented as a percentage of control values obtained from untreated cultures.