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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Todaro GJ, Lazar GK, Green H (1965) The initiation of cell division in a contact-inhibited mammalian cell line. J Cell Physiol 66(3):325–333. https://doi.org/10.1002/jcp.1030660310
Burk RR (1973) A factor from a transformed cell line that affects cell migration. Proc Natl Acad Sci U S A 70(2):369–372. https://doi.org/10.1073/pnas.70.2.369
Al-Lazikani B, Banerji U, Workman P (2012) Combinatorial drug therapy for cancer in the post-genomic era. Nat Biotechnol 30(7):679–692. https://doi.org/10.1038/nbt.2284
Bijnsdorp IV, Giovannetti E, Peters GJ (2011) Analysis of drug interactions. Methods Mol Biol 731:421–434. https://doi.org/10.1007/978-1-61779-080-5_34
Ianevski A, He L, Aittokallio T, Tang J (2017) SynergyFinder: a web application for analyzing drug combination dose-response matrix data. Bioinformatics 33(15):2413–2415. https://doi.org/10.1093/bioinformatics/btx162
Chou T, Martin N (2005) CompuSyn for drug combinations: PC software and user’s guide: a computer program for quantitation of synergism and antagonism in drug combinations, and the determination of IC50 and ED50 and LD50 values. ComboSyn Inc, Paramus, NJ
Kong M, Lee JJ (2008) A semiparametric response surface model for assessing drug interaction. Biometrics 64(2):396–405. https://doi.org/10.1111/j.1541-0420.2007.00882.x
Chou TC (2006) Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 58(3):621–681. https://doi.org/10.1124/pr.58.3.10
Di Veroli GY, Fornari C, Wang D, Mollard S, Bramhall JL, Richards FM, Jodrell DI (2016) Combenefit: an interactive platform for the analysis and visualization of drug combinations. Bioinformatics 32(18):2866–2868. https://doi.org/10.1093/bioinformatics/btw230
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675. https://doi.org/10.1038/nmeth.2089
Poon PY, Yue PY, Wong RN (2017) A device for performing cell migration/wound healing in a 96-well plate. J Vis Exp (121). https://doi.org/10.3791/55411
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kobelt, D., Walther, W., Stein, U.S. (2021). Real-Time Cell Migration Monitoring to Analyze Drug Synergism in the Scratch Assay Using the IncuCyte System. In: Stein, U.S. (eds) Metastasis. Methods in Molecular Biology, vol 2294. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1350-4_9
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1350-4_9
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1349-8
Online ISBN: 978-1-0716-1350-4
eBook Packages: Springer Protocols