Cell migration and invasion assays.pdf

Methods 37 (2005) 208–215 /locate/ymethCell migration and invasion assays Aline Valster a,1, Nhan L. Tran d,1, Mitsutoshi Nakada d, Michael E. Berens d, Amanda Y. Chan a, Marc Symons a,b,c,¤ a Feinstein Institute for Medical Research at North Shore-LIJ, 350 Community Drive, Manhasset, NY 11030, USA b Department of Surgery, North Shore University Hospital, Manhasset, NY 11030, USA c Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA d Neurogenomics Division, The Translational Genomics Research Institute, Phoenix, AZ 85004, USA Accepted 24 May 2005 Abstract The processes of cell migration and invasion are integral to embryonic development and the functioning of adult organisms. Deregulation of these processes contributes to numerous diseases. Ras GTPases and in particular members of the Rho subfamily of GTPases play critical roles in cell migration and invasion. Here, we provide a collection of protocols to assay these functions. We describe two cell migration assays. The monolayer wound healing assay is very easy to implement, whereas the microliter-scale migration assay allows examination of cell behavior on deWned extracellular matrices. We also describe two methods that allow the quantiWcation of tumor cell invasion, a versatile transwell Matrigel invasion assay and an organotypic assay that examines the inva- sion of glioma cells through a rat brain slice. ? 2005 Elsevier Inc. All rights reserved. Keywords: Migration; Invasion; siRNA; Rac; Glioma1. Introduction Cell migration is a process that is critical at many stages of embryonic development, and is essential for tis- sue repair and immune function [1]. Importantly, dereg- ulated motile behavior contributes to pathological processes including tumor angiogenesis and metastasis, atherosclerosis, and arthritis [2–4]. Members of the Ras superfamily of GTPases, most notably the Rho proteins, play a prominent role in cell migration [5]. Thus, the Rac, Cdc42, and