Creation of luminal tissue covered with urothelium by implantation of cultured urothelial cells into the peritoneal cavity.

Abstract

We established the culture condition of seeding urothelial cells onto a scaffold for implantation into the peritoneal cavity and evaluated the histology of implanted urothelial cells. In part 1 of the study cultured porcine bladder urothelial cells were seeded onto 3 types of collagen gel made on microporous membrane, including collagen gel with or without cultured porcine bladder fibroblasts, or a feeder layer. The macroscopic and microscopic appearance of the gel with urothelial cells were examined in vitro. As an in vivo study, cultured porcine bladder urothelial cells were seeded onto a collagen gel/sponge matrix with or without cultured fibroblasts, or a feeder layer. Urothelial cell survival on each matrix was evaluated 28 days after implantation onto the omentum or mesentery of nude rats. In part 2 of the study rat urothelial cells were cultured and seeded onto fibrin gel/atelocollagen sponge matrix as an autologous implantation model. After 7 days of cultivation the matrix was folded with urothelial cells inside, implanted onto the mesentery and serially evaluated. Gel containing cultured fibroblasts was shrunken and basement membrane formation was observed on the gel with cultured fibroblasts or the feeder layer in vitro. Urothelial cells cultured with the feeder layer better survived on the collagen based matrix and formed a hollow-like lumen when implanted into the peritoneal cavity. The regenerated urothelium in an autologous implantation showed the same histological features as normal bladder urothelium. Selection of less degradable matrix and formation of basement membrane are critical for survival of implanted urothelial cells. The regenerated urothelium in an autologous implantation model seems to have the similar properties to the normal urothelium.