title: Multiplex profiling of cellular invasion in 3D cell culture models.
abstract: To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states.
authors.affiliation: Comprehensive Pneumology Center, University Hospital of the Ludwig-Maximilians-University Munich and Helmholtz Zentrum München, Member of the German Center for Lung Research, Munich, Germany. gerald.burgstaller@helmholtz-muenchen.de
authors.forename: Gerald
authors.initials: G
authors.lastname: Burgstaller
authors.forename: Bettina
authors.initials: B
authors.lastname: Oehrle
authors.forename: Ina
authors.initials: I
authors.lastname: Koch
authors.forename: Michael
authors.initials: M
authors.lastname: Lindner
authors.forename: Oliver
authors.initials: O
authors.lastname: Eickelberg
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D008168:Lung / Q000166:cytology; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D019718:Receptors, CXCR4; D004815:Epidermal Growth Factor; D003094:Collagen; D053509:Matrix Metalloproteinase 13