Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates

The overall goal of this set of procedures is to generate three-dimensional cell cultures to function as in vitro breast cancer surrogates, and to present a method for measuring cell growth in histologic sections of the surrogates. Three-dimensional cell culture is a more physiologically relevant method of modelling cell behavior in vitro than is two-dimensional cell culture. By altering cellular composition in experimental conditions, this set of procedures can be adapted to address a variety of biological questions, and to assess the efficacy of candidate therapeutics.

To incorporate the cells into the ECM, on ice, add the cells and the first four related components listed in table one in the accompanying manuscript, into a two millimeter microcentrifuge tube. Next, gently mix them by pipetting, and avoid forming any bubbles. Monitor the pH level using the phenol red in the 10XD MEM to make sure that the mixture shows an orange-pink color, indicating a pH of seven.