In vitro cell migration, invasion, and adhesion assays: from cell imaging to data analysis
Moreno, David F.
Marti, Rosa M.
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Cell migration is a key procedure involved in many biological processes includingembryological development, tissue formation, immune defense or inflammation, andcancer progression. How physical, chemical, and molecularaspects can affect cellmotility is a challenge to understand migratory cells behavior.In vitroassays are excellentapproaches to extrapolate toin vivosituations and study live cells behavior. Here wepresent fourin vitroprotocols that describe step-by-step cell migration, invasion andadhesion strategies and their corresponding image data quantification. These currentprotocols are based ontwo-dimensionalwound healing assays (comparing traditionalpipette tip-scratch assay vs. culture insert assay), 2D individual cell-tracking experimentsby live cell imaging andthree-dimensionalspreading and transwell assays. All together,they cover different phenotypes and hallmarks of cell motility and adhesion, providingorthogonal information that can be used either individually or collectively in many differentexperimental setups. These optimized protocols will facilitate physiological and cellularcharacterization of these processes, which may be used for fast screening of specifictherapeutic cancer drugs for migratory function, novel strategies in cancer diagnosis,and for assaying new molecules involved in adhesion and invasion metastatic propertiesof cancer cells.Keywords: cell migration/invasion, wound healing assay, scratch assay, transwell assay, spreading assay, live cellimaging, data analysis, ImageJ/FijiINTRODUCTIONCell migration is a crucial process where cells must be able tochange and reach their properposition in a given environment to execute their function (te Boekhorst et al., 2016). In multicellularorganisms, this phenomenon plays an important role in gastrulation, embryonic morphogenesis,development of the nervous system, tissue homeostasis, and immune cell trafficking. However,moving cells can be deregulated and contribute to many pathological processes such asinflammation and cancer metastasis (Charras and Sahai, 2014; Mayor and Etienne-Manneville,2016; van Helvert et al., 2018).In cancer development and progression, invasion, and metastasis occurs when tumor cellsdisseminate from the primary tumor spreading through the circulatory and lymphatic systems,invade across the basement membranes and endothelial walls and finally colonize distant organs
Is part ofFrontiers in Cell and Developmental Biology, vol. 7, art. 107
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