Reliable Automated Cytotoxicity Profiling by Combining Multiplexed Cytotoxicity Assays with the CyBio FeliX Application Note · CyBio FeliX

Introduction

In vitro cell-based assays are a convenient and cost-effective way of evaluating test compound toxicity especially in drug discovery research programs. A typical workflow involves titration of the test compound, application to cells, and incubation for several days. We used the CyBio® FeliX (Analytik Jena) instrument as a stand-alone, fully automated system to perform all cytotoxicity assay-specific liquid handling tasks.

Effects of the test compounds were quantified through the application of reagents designed to measure biomarkers of interest. In general terms, a compound or treatment that elicits a cytotoxic effect does so by causing dramatic morphological changes, adversely affecting replication rate or leading to a reduction in viable cell count. Information gained from in vitro methods is an important antecedent to flag potential toxic liabilities. In mechanistic cytotoxicity profiling, unique biomarkers are used to profile the health status of cells. In this report, we used a panel of three assays to assess biomarkers associated with changes in membrane integrity (cytotoxicity), cell number and apoptosis.

When used together, these biomarkers can help distinguish populations of cells that are necrotic, apoptotic or growth arrested.