Three-Dimensional, Quantitative in vitro Assays of Wound Healing Behavior

This chapter describes several assays that allow quantitative evaluation of fundamental aspects of cell behavior involved in the wound healing response in controlled environments with improved physiological relevance. Collagen gels are prepared by neutralizing stock type I collagen solution and raising the temperature to facilitate self-assembly of monomeric collagen into fibrils and forming an entangled network of fibrils with interstitial medium. Disrupting forming gels can affect structural and rheological properties that are important for cell behavior assays. Take care when handling the gels during and after formation. The key is to maintain tight control over cell populations and the cell density used in the assays, as cell behavior can vary widely from passage to passage, and cell density dramatically affects the potential for cell-cell signaling, which, if not accounted for, can cloud results. For each cell, using standard image analysis packages, draw a line segment representing the prevailing orientation of the cell. © 2006