Comparison of a novel redox dye cell growth assay to the ATP bioluminescence assay.

Antisense oligodeoxynucleotides (ASOs) are being studied with increasing frequency to determine their potential role as anticancer drugs. The overwhelming majority of this research continues to be done in vitro. Reliable and accurate determination of cell proliferation and/or inhibition, therefore, is essential for the production of meaningful results. The Alamar Blue assay, a colorimetric/fluorometric redox dye assay, was recently marketed by Alamar Biosciences (Sacramento, CA). In order to determine a potential role for this assay in ASO research, it was extensively tested on ovarian cancer cells in culture under a variety of conditions including uninhibited cell growth, cell kill with cytotoxic chemotherapeutic agents, and growth inhibition with ASOs. Results were compared directly with the ATP bioluminescence assay. We conclude that the Alamar Blue assay correlates poorly with the ATP assay, and it should be used only as a rapid preliminary screening assay. The ATP bioluminescence assay should continue to be considered as the standard in this setting.