title: Generating and characterizing the mechanical properties of cell-derived matrices using atomic force microscopy.
abstract: Mechanical interaction between cells and their surrounding extracellular matrix (ECM) controls key processes such as proliferation, differentiation and motility. For many years, two-dimensional (2D) models were used to better understand the interactions between cells and their surrounding ECM. More recently, variation of the mechanical properties of tissues has been reported to play a major role in physiological and pathological scenarios such as cancer progression. The 3D architecture of the ECM finely tunes cellular behavior to perform physiologically relevant tasks. Technical limitations prevented scientists from obtaining accurate assessment of the mechanical properties of physiologically realistic matrices. There is therefore a need for combining the production of high-quality cell-derived 3D matrices (CDMs) and the characterization of their topographical and mechanical properties. Here, we describe methods that allow to accurately measure the young modulus of matrices produced by various cellular types. In the first part, we will describe and review several protocols for generating CDMs matrices from endothelial, epithelial, fibroblastic, muscle and mesenchymal stem cells. We will discuss tools allowing the characterization of the topographical details as well as of the protein content of such CDMs. In a second part, we will report the methodologies that can be used, based on atomic force microscopy, to accurately evaluate the stiffness properties of the CDMs through the quantification of their young modulus. Altogether, such methodologies allow characterizing the stiffness and topography of matrices deposited by the cells, which is key for the understanding of cellular behavior in physiological conditions.
authors.affiliation: Instituto de Micreoelectrónica de Madrid, CSIC, Isaac Newton 8, 28760, Tres Cantos, Madrid, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Tello
authors.affiliation: Inserm U1109, MN3T, Strasbourg F-67200, France; Université de Strasbourg, Strasbourg F-67000, France; LabEx Medalis, Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Spenlé
authors.affiliation: Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France; Inserm U1121, Strasbourg F-67200, France.
authors.forename: Joseph
authors.initials: J
authors.lastname: Hemmerlé
authors.affiliation: Inserm U1109, MN3T, Strasbourg F-67200, France; Université de Strasbourg, Strasbourg F-67000, France; LabEx Medalis, Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France.
authors.forename: Luc
authors.initials: L
authors.lastname: Mercier
authors.affiliation: Centre d'Immunologie de Marseille - Luminy, Parc Scientifique et Technologique de Luminy, F-13288 Marseille, France.
authors.forename: Roxane
authors.initials: R
authors.lastname: Fabre
authors.affiliation: Inserm U1109, MN3T, Strasbourg F-67200, France; Université de Strasbourg, Strasbourg F-67000, France; LabEx Medalis, Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Allio
authors.affiliation: Inserm U1109, MN3T, Strasbourg F-67200, France; Université de Strasbourg, Strasbourg F-67000, France; LabEx Medalis, Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France. Electronic address: simona@unistra.fr.
authors.forename: Patricia
authors.initials: P
authors.lastname: Simon-Assmann
authors.affiliation: Inserm U1109, MN3T, Strasbourg F-67200, France; Université de Strasbourg, Strasbourg F-67000, France; LabEx Medalis, Université de Strasbourg, Strasbourg F-67000, France; Fédération de Médecine Translationnelle de Strasbourg (FMTS), Strasbourg F-67000, France. Electronic address: jacky.goetz@inserm.fr.
authors.forename: Jacky G
authors.initials: JG
authors.lastname: Goetz
mesh_terms: D000818:Animals; D002417:Cattle; D055119:Elastic Modulus; D005109:Extracellular Matrix / Q000737:chemistry / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology; D051379:Mice; D018625:Microscopy, Atomic Force; D032389:Myocytes, Smooth Muscle / Q000502:physiology; D041681:NIH 3T3 Cells
keywords: Atomic force microscopy; Cell culture; Cell-derived matrices; Extracellular matrix; Topography; Young modulus