Journal ArticleDOI
Sulforhodamine B colorimetric assay for cytotoxicity screening
TLDR
The sulforhodamine B (SRB) assay is used for cell density determination, based on the measurement of cellular protein content, which is an efficient and highly cost-effective method for screening.Abstract:
The sulforhodamine B (SRB) assay is used for cell density determination, based on the measurement of cellular protein content. The method described here has been optimized for the toxicity screening of compounds to adherent cells in a 96-well format. After an incubation period, cell monolayers are fixed with 10% (wt/vol) trichloroacetic acid and stained for 30 min, after which the excess dye is removed by washing repeatedly with 1% (vol/vol) acetic acid. The protein-bound dye is dissolved in 10 mM Tris base solution for OD determination at 510 nm using a microplate reader. The results are linear over a 20-fold range of cell numbers and the sensitivity is comparable to those of fluorometric methods. The method not only allows a large number of samples to be tested within a few days, but also requires only simple equipment and inexpensive reagents. The SRB assay is therefore an efficient and highly cost-effective method for screening.read more
Citations
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Journal ArticleDOI
Neutral red uptake assay for the estimation of cell viability/cytotoxicity.
TL;DR: The neutral red uptake assay provides a quantitative estimation of the number of viable cells in a culture and is cheaper and more sensitive than other cytotoxicity tests (tetrazolium salts, enzyme leakage or protein content).
Journal ArticleDOI
Resistance to therapy caused by intragenic deletion in BRCA2
Stacey L. Edwards,Rachel Brough,Christopher J. Lord,Rachael Natrajan,Radost Vatcheva,Douglas A. Levine,Jeff Boyd,Jorge S. Reis-Filho,Alan Ashworth +8 more
TL;DR: It is shown that resistance to PARP inhibition can be acquired by deletion of a mutation in BRCA2, and new BRCa2 isoforms were expressed in the resistant lines as a result of intragenic deletion of the c.6174delT mutation and restoration of the open reading frame (ORF).
Journal ArticleDOI
Carbon-Dot-Decorated Carbon Nitride Nanoparticles for Enhanced Photodynamic Therapy against Hypoxic Tumor via Water Splitting.
TL;DR: In vitro study showed that PCCN could increase the intracellular O2 concentration and improve the reactive oxygen species generation in both hypoxic and normoxic environments upon light irradiation, and in vivo experiments indicated that P CCN had superior ability to overcome tumor hypoxia.
Journal ArticleDOI
Tumour hypoxia causes DNA hypermethylation by reducing TET activity
Bernard Thienpont,Jessica Steinbacher,Hui Zhao,Flora D’Anna,Anna Kuchnio,Athanasios Ploumakis,Bart Ghesquière,Laurien Van Dyck,Bram Boeckx,Luc Schoonjans,Els Hermans,Frédéric Amant,Vessela N. Kristensen,Kian Peng Koh,Massimiliano Mazzone,Mathew L. Coleman,Thomas Carell,Peter Carmeliet,Diether Lambrechts +18 more
TL;DR: Tumour hypoxia acts as a novel regulator of DNA methylation in tumour suppressor genes by reducing the activity of oxygen-dependent ten-eleven translocation enzymes in human and mouse cells.
Journal ArticleDOI
Haem oxygenase is synthetically lethal with the tumour suppressor fumarate hydratase
Christian Frezza,Liang Zheng,Ori Folger,Kartik N. Rajagopalan,Elaine D. MacKenzie,Livnat Jerby,Massimo Micaroni,Barbara Chaneton,Julie Adam,Ann Hedley,Gabriela Kalna,Ian Tomlinson,Patrick J. Pollard,Dave Watson,Ralph J. DeBerardinis,Tomer Shlomi,Eytan Ruppin,Eyal Gottlieb +17 more
TL;DR: This work predicted and confirmed that targeting this pathway would render Fh1-deficient cells non-viable, while sparing wild-type Fh2-containing cells, and demonstrated that inhibition of haem oxygenation is synthetically lethal when combined with Fh 1 deficiency.
References
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Journal ArticleDOI
New Colorimetric Cytotoxicity Assay for Anticancer-Drug Screening
Philip Skehan,Ritsa Storeng,Dominic A. Scudiero,Anne Monks,James B. McMahon,David T. Vistica,Jonathan T. Warren,Heidi R. Bokesch,Susan Kenney,Michael R. Boyd +9 more
TL;DR: The SRB assay provides a sensitive measure of drug-induced cytotoxicity, is useful in quantitating clonogenicity, and is well suited to high-volume, automated drug screening.
Journal ArticleDOI
Feasibility of a High-Flux Anticancer Drug Screen Using a Diverse Panel of Cultured Human Tumor Cell Lines
Anne Monks,Dominic A. Scudiero,Philip Skehan,Robert H. Shoemaker,Kenneth D. Paull,David T. Vistica,Curtis Hose,John Langley,Paul Cronise,Anne Vaigro-Wolff,Marcia Gray-Goodrich,H. D. Campbell,Joseph G. Mayo,Michael R. Boyd +13 more
TL;DR: A pilot-scale, in vitro, anticancer drug screen utilizing a panel of 60 human tumor cell lines organized into subpanels representing leukemia, melanoma, and cancers of the lung, colon, kidney, ovary, and central nervous system is described.
Journal ArticleDOI
Comparison of In Vitro Anticancer-Drug-Screening Data Generated With a Tetrazolium Assay Versus a Protein Assay Against a Diverse Panel of Human Tumor Cell Lines
Larry Rubinstein,Robert H. Shoemaker,Kenneth D. Paull,Richard M. Simon,S. Tosini,Philip Skehan,Dominic A. Scudiero,Anne Monks,Michael R. Boyd +8 more
TL;DR: A detailed comparison of data generated by each type of assay was undertaken, and results indicate that under the experimental conditions used and within the limits of the data analyses, the assays perform similarly.
Journal Article
Effects of the pH dependence of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan absorption on chemosensitivity determined by a novel tetrazolium-based assay
TL;DR: It is shown that reduction of M TT to MTT-formazan by cells is dependent on the amount of MTT in the incubation medium and can be used to estimate cell numbers in a simple chemosensitivity assay the results of which agree well with a commonly used clonogenic assay.
Journal ArticleDOI
Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing.
Yvonne P. Keepers,Paulo E. Pizao,Godefridus J. Peters,Jannette van Ark-Otte,B. Winograd,B. Winograd,Herbert M. Pinedo +6 more
TL;DR: The sulforhodamine B (SRB) protein stain assay was compared with the tetrazolium (MTT) colorimetric assay for in vitro chemosensitivity testing of various human tumour cell lines and provided a better linearity with cell number and a higher sensitivity.