Journal Article

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

Biermann, Jana, Kovács, Anikó, Karlsson, Per, Larsson, Peter, Forssell-Aronsson, Eva, Parris, Toshima Z., Helou, Khalil, Engqvist, Hanna, Werner Rönnerman, Elisabeth

2020

Overview

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.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/154 / 631/337 / 631/67 / 692/4028 / Antineoplastic Agents - toxicity / Bortezomib / Bortezomib - toxicity / Breast cancer / Cancer / Cancer and Oncology / Cancer och onkologi / Carboplatin / Carboplatin - toxicity / Cell Biology / Cell Survival / Cell viability / Cell viability assay / Cellbiologi / Cisplatin / Cisplatin - toxicity / Dimethyl Sulfoxide - standards / Drug development / drug discovery / Drug Resistance, Neoplasm / Drug screening / Drug Screening Assays, Antitumor - methods / Drug Screening Assays, Antitumor - standards / Evaporation / Experimental design / Humanities and Social Sciences / Humans / Inhibitor drugs / Inhibitory Concentration 50 / MCF-7 Cells / Molecular Biology / Molekylärbiologi / multidisciplinary / oncology / Optimization / Platinum / Proteasome inhibitors / Reproducibility / Reproducibility of Results / Science / Science (multidisciplinary) / Tumor cell lines