Measurement of Cell Migration on Surface-Bound Fibronectin Gradients

Abstract

A novel technique for the quantitative observation of cell migration along linear gradient substrates functionalized with adhesive proteins is presented. Gradients of the cell adhesion molecule fibronectin are generated by the cross diffusion of functionalizable alkanethiols on gold and characterized by X-ray photoelectron spectroscopy and surface plasmon resonance. Two distinct migration assays are described that characterize the movement of either sparsely populated noncontacting cells or a confluent monolayer of cells into free space. The drift speed of bovine aortic endothelial cells is measured and shown to increase along a fibronectin gradient when compared to a uniform control substrate using both assays. The results of these experiments establish reproducible conditions for studies of cell migration on gradients of surface-bound ligands.

Cell adhesion and response to synthetic nanopatterned environments by steering receptor clustering and spatial location

“…Herbert et al (1997) introduced a method for using photoimmobilization of peptides on self-assembled monolayer, while (Wang et al, 2004) merged electrochemical potential gradients with electrosorption of organothiols. Smith et al (2004) produced surface-bound gradients of fibronectin by the crossdiffusion of inert and reactive functionalized alkanethiols on gold. The formation of self-assembled gradients of COOH functional groups allows the covalent link to N-groups of fibronectin.…”

A numerical model for durotaxis

“…Different kinds of cell migration have been modelled using the discrete approach: random motility (Zaman et al, 2005;Dickinson and Tranquillo, 1993), haptotaxis (Smith et al, 2004;Dickinson and Tranquillo, 1993), chemotaxis (Tranquillo and Lauffenburger, 1987;Jabbarzadeh and Abrams, 2005) and galvanotaxis (Schienbein and Gruler, 1993). Concerning durotaxis modelling, to our knowledge, it has been studied only using the continuous approach by Moreo et al (2008).…”

Surface-chemical and -morphological gradients

“…The attachment and movement of cells depends strongly on the surface concentration and the confirmation of adsorbed proteins, and thus it is useful to study this behaviour in a high-throughput manner. Proteins have been covalently coupled to alkanethiol gradients by, for example, an amine group 37,81 or by laser irradiation 78 and they have been ink-jet printed 50 or stamped. 82 Other groups have controlled the adsorption kinetics by changing the contact time of a substrate with a protein solution.…”

“…37 The cell migration and directedness of endothelial cells was also found to increase with a higher laminin density.…”

“…Order-disorder gradients 36 using alkanethiols of different lengths, and wettability gradients have been used to study protein adsorption and cell adhesion. 7,37,38 In 2000, Terrill et al presented a method that relied on the electrochemical desorption of alkanethiols from a fully covered SAM by application of a potential. 39 The width of the potential window and the position of the electrodes thereby determine the width and slope of the gradients (see Fig.…”