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Characterizing the mechanics of cultured cell monolayers

Harris, Andrew R ; Peter, Loic ; Bellis, Julien ; Baum, Buzz ; Kabla, Alexandre J ; Charras, Guillaume T

Proceedings of the National Academy of Sciences - PNAS, 2012-10, Vol.109 (41), p.16449-16454 [Periódico revisado por pares]

United States: National Academy of Sciences

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Título:
Characterizing the mechanics of cultured cell monolayers
Autor: Harris, Andrew R ; Peter, Loic ; Bellis, Julien ; Baum, Buzz ; Kabla, Alexandre J ; Charras, Guillaume T
Assuntos: Actins ; Animals ; Biological Sciences ; Cadherins ; Cadherins - genetics ; Cadherins - metabolism ; Cell Adhesion - physiology ; Cell culture ; Cell Culture Techniques - instrumentation ; Cell Culture Techniques - methods ; Cell Proliferation ; Cells ; Collagen - metabolism ; Collagens ; cultured cells ; Cytoskeleton ; Cytoskeleton - metabolism ; Deformation ; Dogs ; early development ; Embryology ; Epithelial cells ; Fracture mechanics ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; image analysis ; Immunohistochemistry ; Intercellular junctions ; Intercellular Junctions - physiology ; keratin ; Keratins ; Madin Darby Canine Kidney Cells ; Mechanical properties ; microfilaments ; Microscopy, Confocal ; morphogenesis ; myosin ; Physical Sciences ; Research universities ; Stiffness ; Stress, Mechanical ; tissues
É parte de: Proceedings of the National Academy of Sciences - PNAS, 2012-10, Vol.109 (41), p.16449-16454
Notas: http://dx.doi.org/10.1073/pnas.1213301109
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Edited by David A. Weitz, Harvard University, Cambridge, MA, and approved August 22, 2012 (received for review August 1, 2012)
Author contributions: A.R.H., A.J.K., and G.T.C. designed research; A.R.H., J.B., and B.B. performed research; A.R.H., L.P., and G.T.C. contributed new reagents/analytic tools; A.R.H. analyzed data; and A.R.H., A.J.K., and G.T.C. wrote the paper.
Descrição: One-cell-thick monolayers are the simplest tissues in multicellular organisms, yet they fulfill critical roles in development and normal physiology. In early development, embryonic morphogenesis results largely from monolayer rearrangement and deformation due to internally generated forces. Later, monolayers act as physical barriers separating the internal environment from the exterior and must withstand externally applied forces. Though resisting and generating mechanical forces is an essential part of monolayer function, simple experimental methods to characterize monolayer mechanical properties are lacking. Here, we describe a system for tensile testing of freely suspended cultured monolayers that enables the examination of their mechanical behavior at multi-, uni-, and subcellular scales. Using this system, we provide measurements of monolayer elasticity and show that this is two orders of magnitude larger than the elasticity of their isolated cellular components. Monolayers could withstand more than a doubling in length before failing through rupture of intercellular junctions. Measurement of stress at fracture enabled a first estimation of the average force needed to separate cells within truly mature monolayers, approximately ninefold larger than measured in pairs of isolated cells. As in single cells, monolayer mechanical properties were strongly dependent on the integrity of the actin cytoskeleton, myosin, and intercellular adhesions interfacing adjacent cells. High magnification imaging revealed that keratin filaments became progressively stretched during extension, suggesting they participate in monolayer mechanics. This multiscale study of monolayer response to deformation enabled by our device provides the first quantitative investigation of the link between monolayer biology and mechanics.
Editor: United States: National Academy of Sciences
Idioma: Inglês

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Characterizing the mechanics of cultured cell monolayers

Harris, Andrew R ; Peter, Loic ; Bellis, Julien ; Baum, Buzz ; Kabla, Alexandre J ; Charras, Guillaume T

United States: National Academy of Sciences

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