Protocol

Adhesion Protein Protocols

Volume 370 of the series Methods in Molecular Biology™ pp 67-81

Dynamic Assessment of Cell-Matrix Mechanical Interactions in Three-Dimensional Culture

  • W. Matthew PetrollAffiliated withDepartment of Ophthalmology, University of Texas Southwestern Medical Center

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Cell-matrix mechanical interactions play a defining role in a range of biological processes such as developmental morphogenesis and wound healing. Despite current agreement that fibroblasts exert mechanical forces on the extracellular matrix (ECM) to promote structural organization of the collagen architecture, the underlying mechanisms of force generation and transduction to the ECM are not completely understood. Investigation of these processes has been limited, in part, by the technical challenges associated with simultaneous imaging of cell activity and fibrillar collagen organization. To overcome these limitations, we have developed an experimental model in which cells expressing proteins tagged with enhanced green fluorescent protein are plated inside fibrillar collagen matrices, and high magnification time-lapse differential interference contrast and fluorescent imaging is then performed. Using this system, focal adhesion movement and reorganization in isolated cells can be directly correlated with collagen matrix deformation and changes in the mechanical behavior of fibroblasts can be assessed over time.

Key Words

Focal adhesions focal complexes cell motility collagen matrices actomyosin cell mechanics three-dimensional culture