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Kinetic Multi-Spheroid Assays
Multiplexed viability and toxicity measurements enable investigation
of mechanisms of action
In addition to screening for cytotoxicity, live-cell multi-spheroid assays can be used in
conjunction with IncuCyte® Cell Health Reagents to determine the mechanism of action
of drugs by combining data on size with levels of cytotoxicity and apoptosis (Figure 6).
Here, the power of the assay is illustrated, distinguishing the cytotoxic effects of CMP from
that of cycloheximide (CHX). The ability to multiplex reagents with fluorescent-expressing
cell lines provides a depth of information on drug mechanism of action under realistic
physiological conditions.
Figure 6. Cytotoxic and cytostatic mechanisms of action can be differentiated by measuring spheroid size and apoptosis. A549 cells were plated at a density
of 2,000 cells per well and spheroid allowed to form (96 h). Spheroids were treated with increasing concentrations of CMP (top row, 4 nM – 10 μM) or CHX
(bottom row, 4 nM – 10 μM) in the presence of IncuCyte® Annexin V Green reagent (1%). Images were taken every 6 h for 10 d. Time courses show change
in size (Brightfield area) or apoptosis (IncuCyte Annexin V fluorescence intensity) over time. CRCs show the different profiles of cytotoxic and cytostatic
mechanisms.
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