Research for Establishing Bladder Reconstruction with Tissue Engineering

2002 Fiscal Year Final Research Report Summary

• Project/Area Number: 13671629
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Urology
• Research Institution: HOKKAIDO UNIVERSITY
• Principal Investigator: KAKIZAKI Hidehiro Hokkaido Univ. Grad, School of Med., Assi. Pro, 医学部附属病院, 講師 (10241324)
• Co-Investigator(Kenkyū-buntansha): TANAKA Hiroshi Hokkaido Univ. Grad. School of Med., Inst., 大学院・医学研究科, 助手 (60344470) ; SHIBATA Takashi Hokkaido Univ. Grad. School of Med., Inst., 医学部附属病院, 助手 (50292032)
• Project Period (FY): 2001 – 2002
• Keywords: Tissue Engineering / Urothelial cell / Bladder / Reconstruction / Cell culture / Implantation

Research Abstract

Epithelial cells exhibit a variety of morphology depending on the various culture conditions in vitro. Tb establish the best culture condition of seeding urothelial cells onto scaffold, we performed in vitro and in vivo studies. Urothelial cells cultured with feeder layer on collagen gel/collagen sponge matrix can better survive and form a hollow like lumen when implanted into the peritoneal cavity. It is concluded that the selection of less degradable matrix and formation of basement membrane are critical for urothelial cell implantation.
To create a urothelial lining tissue in the rat by implantation of cultured autologous urothelial cells, cultured urothelial cells were seeded onto fibrin gel/atelocollagen sponge matrix. After 7 days of cultivation to attach the cells on the matrix, the matrix was folded with urothelial cells inside, implanted onto the mesenterium of terminal ileum, and then evaluated serially with immunohistochemistry and lectinohistochemistry. The implanted matrix showed macroscopically cystic appearance on the 7th day. Stratified urothelium was observed on the luminal surface and infiltration of stromal cells and microvessels was identified in the matrix. Cystic appearance and urothelial lining remained on the 14th day and the regenerated urothelium showed the same lectinohistochemical staining as normal bladder. The infiltrated stromal cells showed positive immunohistochemical staining to both alpha smooth muscle actin and desmin. Thus, we were successful to create a mimetic hollow organ covered with differentiated urothelium from cultured urothelial cells and fibrin gel/atelocollagen sponge matrix. This technique will contribute to the reconstructive surgery of the urinary tract.