Improving consistency of cell-based assays by using division-arrested cells

Area of Science:

• Biotechnology
• Cell Biology
• Pharmacology

Background:

• Cell-based assays are crucial for high-throughput screening (HTS) in drug discovery.
• The consistency and quality of cell reagents are critical for the robustness of HTS.
• Variability in cell populations can significantly impact assay results and reproducibility.

Purpose of the Study:

• To introduce and validate the use of division-arrested cells for cell-based HTS assays.
• To demonstrate that irreversible cell division arrest can be achieved without compromising cell signaling.
• To compare the performance of division-arrested cells versus growing cells in common HTS platforms.

Main Methods:

• Utilizing mitomycin C to induce irreversible cell division arrest in common cell types.
• Assessing cell viability and potential toxicity post-treatment.
• Evaluating cell signaling properties, including reporter gene assays and calcium flux.
• Comparing assay performance using division-arrested cells versus actively dividing cells.

Main Results:

• Mitomycin C effectively arrested cell division in most tested cell types at non-toxic doses.
• Division-arrested cells maintained key cell signaling properties.
• Division-arrested cells demonstrated favorable performance in reporter and calcium flux assays compared to growing cells.
• The technology allows for decoupling cell production from screening processes.

Conclusions:

• Division-arrested cells provide a stable and reproducible cell reagent for HTS.
• This approach enhances the reliability and efficiency of cell-based screening.
• Division arrest technology facilitates on-demand availability of quality-controlled cell reagents for HTS.