Analysis of ADAMTS Effects on Cell Adhesion and Migration

Abstract

As extracellular enzymes that interact extensively with extracellular matrix (ECM) components, several ADAMTS enzymes are understood to influence aspects of cell adhesion to the ECM and the ability of cells to migrate. A standard approach to investigate the involvement of an ADAMTS in these aspects of mammalian cell behavior involves siRNA-mediated knockdown of the expression of the gene of interest in cell culture, followed by methods for quantification of migratory or adhesive behavior. We describe here two methods for cell migration quantification: a time-lapse videomicroscopy method suitable for measuring single cell migration in sparse cultures that allows for determination of migration speed and directionality (persistence), and scratch wound assays for directional migration in confluent cell monolayers. We also present assays to quantify total adhesion to ECM components, as well as more detailed visualization and quantification of focal adhesion structures.