For whom the cells pull: Hydrogel and micropost devices for measuring traction forces.
While performing several functions, adherent cells deform their surrounding substrate via stable adhesions that connect the intracellular cytoskeleton to the extracellular matrix. The traction forces that deform the substrate are studied in mechanotrasduction because they are affected by the mechanics of the extracellular milieu. We review the development and application of two methods widely used to measure traction forces generated by cells on 2D substrates: (i) traction force microscopy with polyacrylamide hydrogels and (ii) calculation of traction forces with arrays of deformable microposts. Measuring forces with these methods relies on measuring substrate displacements and converting them into forces. We describe approaches to determine force from displacements and elaborate on the necessary experimental conditions for this type of analysis. We emphasize device fabrication, mechanical calibration of substrates and covalent attachment of extracellular matrix proteins to substrates as key features in the design of experiments to measure cell traction forces with polyacrylamide hydrogels or microposts. We also report the challenges and achievements in integrating these methods with platforms for the mechanical stimulation of adherent cells. The approaches described here will enable new studies to understand cell mechanical outputs as a function of mechanical inputs and advance the understanding of mechanotransduction mechanisms.
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, United States; Stanford Cardiovascular Institute, Stanford University, Stanford, CA 94305, United States.
Alexandre J S
AJ
Ribeiro
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, United States; Stanford Bioengineering, Stanford University, Stanford, CA 94305, United States.
Aleksandra K
AK
Denisin
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, United States.
Robin E
RE
Wilson
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, United States; Stanford Cardiovascular Institute, Stanford University, Stanford, CA 94305, United States; Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, United States. Electronic address: pruitt@stanford.edu.
Beth L
BL
Pruitt
D000818:Animals; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media; D055119:Elastic Modulus; D020724:Elastomers; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods
D015815:Cell Adhesion Molecules; D003470:Culture Media; D020724:Elastomers; D020100:Hydrogels; D011108:Polymers; C036925:elastomeric polymer
Displacements; Mechanical calibration; PDMS microposts; Polyacrylamide hydrogels; Surface functionalization; Traction force microscopy