Extensive tracheal defects after intensive care medicine, trauma, or large resections in tumor surgery remain a major challenge in plastic and reconstructive surgery. Defects that cannot be satisfactorily treated by complicated and costly reconstructive techniques reveal a need for an alloplastic tracheal replacement. Recent experimental and clinical studies in the development of alloplastic tracheal prostheses proved that the lack of an epithelial lining on the luminal surfaces and inadequate biophysical properties and shapes of the prostheses were the main causes for failure of these prostheses. In this study a cell-seeding technique has been used. Adhesion, spreading, and differentiation of seeded mucosa cells on biomaterials in vitro were observed by scanning electron microscopy (SEM). Chemical properties and surface structure of the material influenced the differentiation process. Epithelium formation of incorporated tracheal prostheses was tested in animal experiments. Isolated respiratory cells were seeded into implanted tubular prostheses of porous polyurethane or expanded polytetrafluorethylene. Light microscopy and SEM showed the tendency of epithelium formation on the surface of the lumen. Vigorous cell layers, predominantly as multiple cell layers of squamous epithelium, were observed. Ciliated or mucus cells were not detected. It can be stated that the epithelium formation on incorporated porous implants is possible. Further studies of the stability and the differentiation process of the epithelium on such implants is needed before an introduction of tracheal replacements into the clinical practice can be considered.