The development of new bladder augmentation technique using cultured bladder epithelial cell sheets.

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Bladder reconstruction using gastrointestinal flaps may induce severe complications. The majority of the complications are related to intestinal mucosa. We have hypothesized that the replacement of gastrointestinal mucosa with urothelial cells could reduce clinical complications. The purpose of this study was to examine whether intact urothelial cell sheets harvested from new temperature-responsive cell culture systems without proteolytic enzyme treatment (1) would be autografted successfully onto demucosalized gastric flaps in a dog model to promote improved regeneration of native-like urothelium. Viable urothelium was obtained from beagle dogs by partial cystectomy (n=8). Harvested urothelial cells were seeded on temperature-responsive culture dishes modified with the thermally sensitive polymer, poly(N-isopropylacrylamide). Urothelial cells cultured for three weeks generated urothelial cell sheets that were non-invasively harvested without enzymatic treatment from these dishes simpl … More y by reducing culture temperature. Urothelial cell sheets were autografted onto surgically demucosalized gastric flaps. Three weeks after autografting, dogs were sacrificed and the gastric flaps with the urothelial cell sheets were examined. Cell and tissue characteristics were compared between these urothelial cell sheet-grafted gastric flaps and native urothelium. Ultrafine structures were also examined. Intact urothelial cell sheets harvested from temperature-responsive surfaces by reducing culture temperature without any proteolytic enzymes exhibit unique properties for transplantation. Five of the eight urothelial cell sheet-grafted flaps showed viable urothelial regeneration. Urothelial cell sheets attached spontaneously to demucosalized tissue surfaces completely without any suture or fixing, and developed into a stratified viable epithelium highly similar to native urothelium. Regenerated urothelium remained unstained by anti-proton pump antibody, which typically stains epithelial cells positively in gastric mucosal layers. Urothelial cell sheets were autografted onto dog demucosalized gastric flaps successfully without suture or fixation requirements, generating a multilayered urothelium in vivo. The intact cell sheet grafting method rapidly produces native-like epithelium in vivo. This versatile technology should prove useful in urinary tract tissue engineering and surgical reconstructions. Less