Bioartificial mucosa can be fabricated with living cells in vitro and used for mucosal grafting. In this report, we describe our culturing methods for preparation of bioartificial mucosa and its morphological characteristics. Bioartificial mucosa has bilayered components, namely, mucosal epithelium and submucosal tissue. Submucosal tissue was composed of fibroblasts and type I collagen lattice. Mucosal epithelium was formed by an epithelial segment grafted on the submucosal tissue surface. Between epithelial and submucosal tissues, basement membrane was observed under a transmission electron microscope. In animal experiments, the bioartificial mucosa was well vascularized and survived completely without immunological rejections. Based on the findings from these animal experiments, we applied this bioartificial mucosa to humans and succeeded in reconstructing their mucosal defects. We also present an overview of the problems relevant to the use of such methods.