Three-Dimensional Organotypic Culture of Stratified Epithelia

One of the limitations of conventional tissue culture on flat two-dimensional surfaces is the loss of complex interactions between the epithelium and stroma. We have devised a culture system that recreates the salient features of the stratified epithelium using primary cell cultures from mouse models. The protocol described here is applicable to the esophageal epithelium, but stratified epithelial cells from other organs (e.g., skin) can be grown. Once established, the system can be used to interrogate the effect of various pharmacologic and genetic manipulations on epithelial homeostasis and invasion.