Research for Establishing Bladder Reconstruction with Tissue Engineering

利用组织工程建立膀胱重建的研究

• 批准号: 13671629
• 负责人: KAKIZAKI Hidehiro
• 金额: $ 2.24万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671629/
• 关键词: Tissue Engineering; Urothelial cell; Bladder; Reconstruction; Cell culture; Implantation

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.

上皮细胞在体外不同的培养条件下表现出不同的形态。通过体外和体内实验，确定了体外培养尿路上皮细胞的最佳条件。在胶原凝胶/胶原海绵基质上培养有饲养层的尿路上皮细胞，植入腹腔后能更好地存活并形成空心样管腔。因此，选择不易降解的基质和基底膜的形成对尿路上皮细胞的植入至关重要。将培养的自体尿路上皮细胞植入纤维蛋白凝胶/间胶原海绵基质，制备大鼠尿路上皮内膜组织。培养7 d后，将细胞贴附于基质上，将基质折叠，其中包覆尿路上皮细胞，植入回肠末端肠系膜，进行免疫组织化学和凝集素组织化学连续评价。植入的基质在第7天出现宏观囊状外观。管腔表面可见层状尿路上皮，基质中可见基质细胞和微血管浸润。第14天仍有囊性外观和尿路上皮，再生的尿路上皮显示与正常膀胱相同的凝集素组织化学染色。浸润间质细胞对α -平滑肌肌动蛋白和desmin免疫组化染色均呈阳性。因此，我们成功地从培养的尿路上皮细胞和纤维蛋白凝胶/间胶原海绵基质中创建了覆盖分化尿路上皮的模拟中空器官。这项技术将有助于泌尿道的重建手术。