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Optimization of cell viability assays to improve replicability and reproducibility of cancer drug sensitivity screens.

Journal article
Authors Peter Larsson
Hanna Engqvist
Jana Biermann
Elisabeth Werner Rönnerman
Eva Forssell-Aronsson
Anikó Kovács
Per Karlsson
Khalil Helou
Toshima Z Parris
Published in Scientific Reports
Volume 10
Issue 1
ISSN 2045-2322
Publication year 2020
Published at Institute of Clinical Sciences, Department of Radiation Physics
Institute of Clinical Sciences, Department of Oncology
Sahlgrenska Cancer Center
Language en
Keywords Cell viability assay, oncology, drug discovery, drug screening
Subject categories Cell and molecular biology, Cell Biology, Cancer and Oncology


Cancer drug development has been riddled with high attrition rates, in part, due to poor reproducibility of preclinical models for drug discovery. Poor experimental design and lack of scientific transparency may cause experimental biases that in turn affect data quality, robustness and reproducibility. Here, we pinpoint sources of experimental variability in conventional 2D cell-based cancer drug screens to determine the effect of confounders on cell viability for MCF7 and HCC38 breast cancer cell lines treated with platinum agents (cisplatin and carboplatin) and a proteasome inhibitor (bortezomib). Variance component analysis demonstrated that variations in cell viability were primarily associated with the choice of pharmaceutical drug and cell line, and less likely to be due to the type of growth medium or assay incubation time. Furthermore, careful consideration should be given to different methods of storing diluted pharmaceutical drugs and use of DMSO controls due to the potential risk of evaporation and the subsequent effect on dose-response curves. Optimization of experimental parameters not only improved data quality substantially but also resulted in reproducible results for bortezomib- and cisplatin-treated HCC38, MCF7, MCF-10A, and MDA-MB-436 cells. Taken together, these findings indicate that replicability (the same analyst re-performs the same experiment multiple times) and reproducibility (different analysts perform the same experiment using different experimental conditions) for cell-based drug screens can be improved by identifying potential confounders and subsequent optimization of experimental parameters for each cell line.

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