title: The IsoStretcher: An isotropic cell stretch device to study mechanical biosensor pathways in living cells.
abstract: Mechanosensation in many organs (e.g. lungs, heart, gut) is mediated by biosensors (like mechanosensitive ion channels), which convert mechanical stimuli into electrical and/or biochemical signals. To study those pathways, technical devices are needed that apply strain profiles to cells, and ideally allow simultaneous live-cell microscopy analysis. Strain profiles in organs can be complex and multiaxial, e.g. in hollow organs. Most devices in mechanobiology apply longitudinal uniaxial stretch to adhered cells using elastomeric membranes to study mechanical biosensors. Recent approaches in biomedical engineering have employed intelligent systems to apply biaxial or multiaxial stretch to cells. Here, we present an isotropic cell stretch system (IsoStretcher) that overcomes some previous limitations. Our system uses a rotational swivel mechanism that translates into a radial displacement of hooks attached to small circular silicone membranes. Isotropicity and focus stability are demonstrated with fluorescent beads, and transmission efficiency of elastomer membrane stretch to cellular area change in HeLa/HEK cells. Applying our system to lamin-A overexpressing fibrosarcoma cells, we found a markedly reduced stretch of cell area, indicative of a stiffer cytoskeleton. We also investigated stretch-activated Ca(2+) entry into atrial HL-1 myocytes. 10% isotropic stretch induced robust oscillating increases in intracellular Fluo-4 Ca(2+) fluorescence. Store-operated Ca(2+) entry was not detected in these cells. The Isostretcher provides a useful versatile tool for mechanobiology.
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Schürmann
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany; Department of Physics, Biophysics Group, FAU, Henkestr. 91, 91052 Erlangen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Wagner
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Herlitze
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Fischer
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Gumbrecht
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Wirth-Hücking
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Prölß
authors.affiliation: Department of Physics, Biophysics Group, FAU, Henkestr. 91, 91052 Erlangen, Germany.
authors.forename: L A
authors.initials: LA
authors.lastname: Lautscham
authors.affiliation: Department of Physics, Biophysics Group, FAU, Henkestr. 91, 91052 Erlangen, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Fabry
authors.affiliation: Department of Physics, Biophysics Group, FAU, Henkestr. 91, 91052 Erlangen, Germany.
authors.forename: W H
authors.initials: WH
authors.lastname: Goldmann
authors.affiliation: Molecular Cardiology Division, Victor Chang Cardiac Research Institute, 405 Liverpool St, Darlinghurst, NSW 2010 Sydney, Australia.
authors.forename: V
authors.initials: V
authors.lastname: Nikolova-Krstevski
authors.affiliation: Mechanosensory Biophysics Laboratory, Victor Chang Cardiac Research Institute, Darlinghurst NSW 2010, Australia; St Vincent's Clinical School, University of New South Wales, Darlinghurst NSW 2010, Australia.
authors.forename: B
authors.initials: B
authors.lastname: Martinac
authors.affiliation: Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Paul-Gordan-Str.3, 91052 Erlangen, Germany. Electronic address: oliver.friedrich@mbt.uni-erlangen.de.
authors.forename: O
authors.initials: O
authors.lastname: Friedrich
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004867:Equipment Design; D005354:Fibrosarcoma / Q000473:pathology; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D040542:Mechanotransduction, Cellular; D008567:Membranes, Artificial; D032383:Myocytes, Cardiac / Q000378:metabolism; D013314:Stress, Mechanical
chemical_list: D004129:Dimethylpolysiloxanes; D008567:Membranes, Artificial; D002118:Calcium
keywords: Cell stiffness; Cell stretch device; Elastomer membrane; Isotropic stretch; Mechanobiology; Mechanosensor