title: Evaluation of viability assays for anthocyanins in cultured cells.
abstract: The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is a widely accepted cytotoxicity assay which can produce inaccurate results due to possible interference with the antioxidant property of anthocyanins. Alternative methods to the MTT assay, such as BrdU (DNA-based) and CellTiter-Glo (ATP-based) assays were evaluated to assess anthocyanin cytotoxicity, derived from blackberry in LNCaP, MCF-7 and MDA-MB-453 cell lines. The standard cell counting method was the reference assay. Greater correlation of cell viability values following anthocyanin exposure was obtained from multiple cell lines with the alternative assays when compared with cell counting. MTT and cell counting results were not always correlated, albeit this was a function of cell type. In particular, poor correlations between cell counting and MTT procedures used to assess cytotoxicity of anthocyanins were observed in the MDA-MB-453 cell lines. Comparison of cytotoxicity derived from alternative assays and the MTT assays with the cell counting method was dependent on the assay procedure and the cell type. The LC(50) of blackberry crude extract ranged from 0.4 to 9.4 mg/mL between assays and across all cell lines, whereas a semi-purified anthocyanin extract was not cytotoxic. Cytotoxicity evaluation of polyphenolic-rich extracts using BrdU and CellTiter-Glo assays as alternatives to the MTT method is recommended.
authors.affiliation: Food, Nutrition, and Health, Faculty of Land and Food Systems, University of British Columbia, 2205 East Mall, Vancouver, B.C., Canada V6T1Z4. ingrid.elisia@gmail.com
authors.forename: Ingrid
authors.initials: I
authors.lastname: Elisia
authors.forename: David G
authors.initials: DG
authors.lastname: Popovich
authors.forename: Chun
authors.initials: C
authors.lastname: Hu
authors.forename: David D
authors.initials: DD
authors.lastname: Kitts
mesh_terms: D000872:Anthocyanins / Q000032:analysis* / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000032:analysis* / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000188:drug therapy; D002452:Cell Count; D045744:Cell Line, Tumor; D002470:Cell Survival; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D005260:Female; D005638:Fruit / Q000737:chemistry*; D006801:Humans; D008297:Male; D010936:Plant Extracts / Q000032:analysis / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000188:drug therapy
chemical_list: D000872:Anthocyanins; D000970:Antineoplastic Agents; D010936:Plant Extracts