title: Functional screening with a live cell imaging-based random cell migration assay.
abstract: Cell migration, essential in cancer progression, is a complex process comprising a number of spatiotemporally regulated and well-coordinated mechanisms. In order to study (random) cell migration in the context of responses to various external cues (such as growth factors) or intrinsic cell signaling, a number of different tools and approaches have been developed. In order to unravel the key pathways and players involved in the regulation of (cancer) cell migration, a systematical mapping of the players/pathways is required. For this purpose, we developed a cell migration assay based on automatic high-throughput microscopy screen. This approach allows for screening of hundreds of genes, e.g., those encoding various kinases and phosphatases but can also be used for screening of drugs libraries. Moreover, we have developed an automatic analysis pipeline comprising of (a) automatic data acquisition (movie) and (b) automatic analysis of the acquired movies of the migrating cells. Here, we describe various facets of this approach. Since cell migration is essential in progression of cancer metastasis, we describe two examples of experiments performed on highly motile (metastatic) cancer cells.
authors: Leiden/Amsterdam Center for Drug Research, Division of Toxicology, University of Leiden, Leiden, The Netherlands.
authors: Wies
authors: W
authors: van Roosmalen
authors: Sylvia E
authors: SE
authors: Le Dévédec
authors: Sandra
authors: S
authors: Zovko
authors: Hans
authors: H
authors: de Bont
authors: Bob
authors: B
authors: van de Water
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D054443:Cell Migration Assays / Q000379:methods*; D002465:Cell Movement; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014584:User-Computer Interface
chemical_list: D003470:Culture Media; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004815:Epidermal Growth Factor; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human