Evaluation of division-arrested cells for cell-based high-throughput screening and profiling

Area of Science:

• Biochemistry
• Cell Biology
• Pharmacology

Background:

• Cell-based high-throughput screening (HTS) faces challenges with just-in-time cell supply.
• Variability in cell quality (passage effect, cell cycle, confluency) impacts HTS assay consistency.
• Cell division-arrest and cryopreservation offer solutions for assay-ready cell reagents.

Purpose of the Study:

• To compare the performance of division-arrested versus dividing cells in cell-based assays.
• To evaluate the utility of division-arrested cells as assay-ready reagents for HTS.
• To assess the impact of cell division status on assay variability and data reproducibility.

Main Methods:

• Two assay types were used: a receptor tyrosine kinase assay (A431 cells) and a secretion reporter assay (secreted alkaline phosphatase).
• Assays were conducted using both dividing and division-arrested cells.
• Compound potency (IC50) and rank order were compared between the two cell types.

Main Results:

• Division-arrested and dividing cells produced comparable basal and maximal signals at equivalent cell densities.
• Similar IC50 values for reference inhibitors were observed in both cell types across both assays.
• The rank order of compound potency was highly consistent between division-arrested and dividing cells in the secretion reporter assay.
• Division-arrested cells demonstrated reduced data variability under specific assay conditions.

Conclusions:

• Division-arrested cells can effectively substitute for dividing cells in many cell-based assays.
• Utilizing division-arrested cells offers potential advantages in terms of consistency and reduced variability for HTS.
• Cell division-arrest technology enhances the reliability of cell-based profiling and structure-activity relationship studies.