title: Quantitative evaluation of single cell spread on collagen matrices.
abstract: Cells change their morphology as a response to environmental cues. The quantitative evaluation of single cell spread on extracellular matrices, such as type I collagen, is a key tool in cancer research. Inherent to the manual scoring of cellular spread is inter-observer but also intra-observer variation. To overcome these problems, we have developed the Morphology Analysis Software (MAS). MAS scores phase-contrast images of cells on native type I collagen gels and identifies whether a cell has a spread or round morphology using a combination of four unique parameters: the presence of a cellular extension, the cell area, the cell eccentricity and cell circularity. The MAS software scores are equivalent to the average score of five independent observers but MAS is faster, more objective and standardized. A functional screening assay using six cytokines identified TGFα as a stimulator of HCT8/E11 and SK-BR-3 single cell spreading on top of type I collagen gels. This change in morphology correlates with increased migration potential as evidenced by xCELLigence migration assays and are counteracted by EGFR signaling pathway inhibitors. This underscores the use of morphology classification on a population of unlabeled cells as read-out of an important cancer cell property and the potential for the MAS software in drug screening strategies.
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: De Vlieghere
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Wagemans
authors.affiliation: DCILabs, Keerbergen, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: De Backer
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: Z
authors.initials: Z
authors.lastname: Drebert
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Tommelein
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: Q
authors.initials: Q
authors.lastname: Rousseau
authors.affiliation: DCILabs, Keerbergen, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Weyn
authors.affiliation: Cancer Research Institute Ghent (CRIG), Ghent University, Belgium; Department of Biochemistry, Fac. Medicine, Gent University, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Van Troys
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Bracke
authors.affiliation: Laboratory of Experimental Cancer Research, Ghent University, Belgium; Cancer Research Institute Ghent (CRIG), Ghent University, Belgium. Electronic address: olivier.dewever@ugent.be.
authors.forename: O
authors.initials: O
authors.lastname: De Wever
mesh_terms: D001331:Automation; D002452:Cell Count; D045744:Cell Line, Tumor; D054443:Cell Migration Assays / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects*; D048430:Cell Shape / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D059010:Single-Cell Analysis / Q000379:methods*; D012984:Software; D016211:Transforming Growth Factor alpha / Q000494:pharmacology
chemical_list: D016207:Cytokines; D047428:Protein Kinase Inhibitors; D016211:Transforming Growth Factor alpha; D003094:Collagen
keywords: Cell morphology; Migration; Native type I collagen; Phase-contrast