New Colorimetric Cytotoxicity Assay for Anticancer-Drug Screening

作者：Philip Skehan, Ritsa Storeng, Dominic A. Scudiero, Anne P. Monks, jeannette mcmahon, David Thomas Vistica, James T. Warren, Heidi Rose Bokesch, Susan Kenney, Michael R. Boyd · 发表于：JNCI Journal of the National Cancer Institute · 年份：1990 · DOI：10.1093/jnci/82.13.1107 · 被引用次数：9828 · 研究领域：3D Printing in Biomedical Research、bioluminescence and chemiluminescence research、Viral Infectious Diseases and Gene Expression in Insects

We have developed a rapid, sensitive, and inexpensive method for measuring the cellular protein content of adherent and suspension cultures in 96-well microtiter plates. The method is suitable for ordinary laboratory purposes and for very large-scale applications, such as the National Cancer Institute's disease-oriented in vitro anticancer-drug discovery screen, which requires the use of several million culture wells per year. Cultures fixed with trichloroacetic acid were stained for 30 minutes with 0.4% (wt/vol) sulforhodamine B (SRB) dissolved in 1% acetic acid. Unbound dye was removed by four washes with 1% acetic acid, and protein-bound dye was extracted with 10 mM unbuffered Tris base [tris (hydroxymethyl)aminomethane] for determination of optical density in a computer-interfaced, 96-well microtiter plate reader. The SRB assay results were linear with the number of cells and with values for cellular protein measured by both the Lowry and Bradford assays at densities ranging from sparse subconfluence to multilayered supraconfluence. The signal-to-noise ratio at 564 nm was approximately 1.5 with 1,000 cells per well. The sensitivity of the SRB assay compared favorably with sensitivities of several fluorescence assays and was superior to those of both the Lowry and Bradford assays and to those of 20 other visible dyes. The SRB assay provides a colorimetric end point that is nondestructive, indefinitely stable, and visible to the naked eye. It provides a sensitive measure of...