title: Microfluidic Migration and Wound Healing Assay Based on Mechanically Induced Injuries of Defined and Highly Reproducible Areas.
abstract: All cell migration and wound healing assays are based on the inherent ability of adherent cells to move into adjacent cell-free areas, thus providing information on cell culture viability, cellular mechanisms and multicellular movements. Despite their widespread use for toxicological screening, biomedical research and pharmaceutical studies, to date no satisfactory technological solutions are available for the automated, miniaturized and integrated induction of defined wound areas. To bridge this technological gap, we have developed a lab-on-a-chip capable of mechanically inducing circular cell-free areas within confluent cell layers. The microdevices were fabricated using off-stoichiometric thiol-ene-epoxy (OSTEMER) polymer resulting in hard-polymer devices that are robust, cost-effective and disposable. We show that the pneumatically controlled membrane deflection/compression method not only generates highly reproducible (RSD 4%) injuries but also allows for repeated wounding in microfluidic environments. Performance analysis demonstrated that applied surface coating remains intact even after multiple wounding, while cell debris is simultaneously removed using laminar flow conditions. Furthermore, only a few injured cells were found along the edge of the circular cell-free areas, thus allowing reliable and reproducible cell migration of a wide range of surface sensitive anchorage dependent cell types. Practical application is demonstrated by investigating healing progression and endothelial cell migration in the absence and presence of an inflammatory cytokine (TNF-α) and a well-known cell proliferation inhibitor (mitomycin-C).
authors.affiliation: BioSensor Technologies, AIT Austrian Institute of Technology GmbH , Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Drago
authors.initials: D
authors.lastname: Sticker
authors.affiliation: BioSensor Technologies, AIT Austrian Institute of Technology GmbH , Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Sarah
authors.initials: S
authors.lastname: Lechner
authors.affiliation: Bioinformatics and High Performance Computing, Austrian Centre of Industrial Biotechnology , Muthgasse 11, 1190 Vienna, Austria.|TGM-Technologisches Gewerbemuseum , Wexstraße 19-23, 1200 Vienna, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Jungreuthmayer
authors.affiliation: Bioinformatics and High Performance Computing, Austrian Centre of Industrial Biotechnology , Muthgasse 11, 1190 Vienna, Austria.|Department of Biotechnology, University of Natural Resources and Life Sciences , Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Zanghellini
authors.affiliation: Faculty of Technical Chemistry, Vienna University of Technology , Getreidemarkt 9, 1060 Vienna, Austria.
authors.forename: Peter
authors.identifier: 0000-0002-7625-2445
authors.initials: P
authors.lastname: Ertl
mesh_terms: D002465:Cell Movement / Q000187:drug effects; D004867:Equipment Design; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D016685:Mitomycin / Q000494:pharmacology; D059008:Time-Lapse Imaging; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014945:Wound Healing / Q000187:drug effects
chemical_list: D014409:Tumor Necrosis Factor-alpha; D016685:Mitomycin