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Real-time cell migration monitoring to analyze drug synergism in the scratch assay using the IncuCyte system

Item Type:Article
Title:Real-time cell migration monitoring to analyze drug synergism in the scratch assay using the IncuCyte system
Creators Name:Kobelt, D. and Walther, W. and Stein, U.S.
Abstract:Drug-mediated interference with metastasis represents a key approach to improve cancer therapy. In this regard, appropriate in vitro assays are needed to identify drugs, which inhibit cell migration as one feature for metastatic potential of cancer cells. One such migration assay is the wound healing or scratch assay, designed to allow cells for closure of an artificially generated gap (wound/scratch) in the monolayer. To identify possibly effective anti-migratory drugs as monotherapy or as synergistic drug combination, novel screening tools besides viability measurements at the experimental endpoint are needed. In this context, particularly drug combinations allow to increase treatment efficacy paralleled by lowered side effects. Here, a protocol for real-time monitoring cellular motility and its inhibition by anti-migratory drugs and combinations by the IncuCyte system and a 96-well scratch assay is described. A pipetting scheme allowing data collection for synergy calculation using one plate per replicate is provided. Using the IncuCyte System 2, drug combinations built of three biological replicates each using three technical replicates can be tested in parallel within hours to few days to accelerate identification of efficient antimetastatic drugs.
Keywords:Drug Interaction, Synergy, IncuCyte, Combenefit, Scratch Assay, Cell Motility, Animals
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Metastasis : methods and protocols
Publisher:Springer / Humana Press
Page Range:133-142
Official Publication:https://doi.org/10.1007/978-1-0716-1350-4_9
PubMed:View item in PubMed
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