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Evaluation of viability assays for anthocyanins in cultured cells
Author(s) -
Elisia Ingrid,
Popovich David G.,
Hu Chun,
Kitts David D.
Publication year - 2008
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.1069
Subject(s) - cytotoxicity , mtt assay , chemistry , viability assay , anthocyanin , cell culture , cell growth , cell counting , microbiology and biotechnology , cell , biochemistry , in vitro , biology , cell cycle , food science , genetics
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. Copyright © 2008 John Wiley & Sons, Ltd.