Single Cell Durotaxis Assay for Assessing Mechanical Control of Cellular Movement and Related Signaling Events
| Autor: | Nyla Naim, Kathryn V Svec, Alan K. Howe, Johnathan B Patterson |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
General Chemical Engineering
Biosensing Techniques Traction force microscopy Fluorescence Article General Biochemistry Genetics and Molecular Biology Focal adhesion Extracellular matrix 03 medical and health sciences 0302 clinical medicine Stress Physiological Cell Line Tumor medicine Animals Humans Mechanotransduction 030304 developmental biology 0303 health sciences Migration Assay Durotaxis General Immunology and Microbiology Chemotaxis General Neuroscience Stiffness Hydrogels Fibroblasts Microspheres Rats Self-healing hydrogels Biophysics Single-Cell Analysis medicine.symptom 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| Popis: | Durotaxis is the process by which cells sense and respond to gradients of tension. In order to study this process in vitro, the stiffness of the substrate underlying a cell must be manipulated. While hydrogels with graded stiffness and long-term migration assays have proven useful in durotaxis studies, immediate, acute responses to local changes in substrate tension allow focused study of individual cell movements and subcellular signaling events. To repeatably test the ability of cells to sense and respond to the underlying substrate stiffness, a modified method for application of acute gradients of increased tension to individual cells cultured on deformable hydrogels is used which allows for real time manipulation of the strength and direction of stiffness gradients imparted upon cells in question. Additionally, by fine tuning the details and parameters of the assay, such as the shape and dimensions of the micropipette or the relative position, placement, and direction of the applied gradient, the assay can be optimized for the study of any mechanically sensitive cell type and system. These parameters can be altered to reliably change the applied stimulus and expand the functionality and versatility of the assay. This method allows examination of both long term durotactic movement as well as more immediate changes in cellular signaling and morphological dynamics in response to changing stiffness. |
| Databáze: | OpenAIRE |
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