Reconstitution of stratified murine and human oesophageal epithelia in anin vivotransplant culture system

OBJECTIVE: The molecular and cellular events responsible for regulating development of the oesophageal epithelium are not well understood. At least in part, this is due to the lack of a suitable model system with which to study the process. Here, we report development of a manipulable in vivo transplant model for mouse or human oesophageal epithelium. MATERIAL AND METHODS: Epithelial cells were isolated from mouse or human oesophagus and inoculated into de-epithelialized and devitalized rat tracheas. The rat trachea, containing cells, was placed subcutaneously under the dorsal skin of immunodeficient mice. RESULTS: We show that a multilayered stratified squamous epithelium can be generated in 4-6 weeks from as few as 5 x 10(4) isolated oesophageal epithelial cells. The reconstituted epithelium recapitulates many of the structural and histological features of the normal oesophageal epithelium, including a basal layer of cuboidal-like cells, suprabasal layers of differentiating squamous cells and, in the case of murine cells, a superficial layer of cornified material. CONCLUSION: Our model can be used to generate a multilayered normal murine or human epithelium from a single cell suspension of oesophageal epithelial cells. The ability to genetically manipulate the cells prior to growth in the model is a powerful tool with which to study the molecular mechanisms involved in the development of normal oesophagus or in pathogenic processes such as Barrett's metaplasia or tumorigenesis.