Localized application of mechanical and biochemical stimuli in 3-D culture

Abstract

The goal of this study was to investigate the responses of isolated cells in 3-D culture to localized application of mechanical and biochemical signals. Corneal fibroblasts were plated inside collagen matrices for 24 hours, then imaged using time-lapse DIC. For mechanical perturbation, a microinjection needle (Femtotip) was inserted axially into the ECM, then displaced laterally to alter local ECM stress. For biochemical stimulation, PDGF or vehicle control solution was microinjected into the matrix. Compressing the ECM perpendicular to the cell axis had no appreciable effect on cell behavior. However, pushing the ECM parallel to the cell axis induced rapid cellular contraction, followed by secondary cell spreading and tractional force generation. Injection of PDGF induced a similar cell spreading response. Cells in 3-D matrices showed remarkable plasticity, and extension of pseudopodia could be induced at both the leading and trailing edges of migrating cells. © 2008 Wiley-Liss, Inc.