Han Ping, Luo Jing-Cong, Zhi Wei, Tan Bo, Li Xiu-Qun, Yang Zhi-Ming
Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 May;39(3):481-4, 510.
To develop a method for reconstructing urothelial structures using tissue engineering techniques.
The bladder mucosa of a puppy dog was mechanically separated, cutted into pieces, and digested with trypsin. The obtained bladder transitional epithelia were cultured with DKSFM and then transplanted to the surface of the small intestinal submucosa (SIS) framework. The compound of the epithelia and small intestinal submucosa (SIS) materials were planted under the back skin of the nude rats. The rats were sacrificed at week 1, 2, 4 and 8 after the transplant. Histological and immunohistochemical examinations were performed to the implanted samples.
The bladder transitional epithelia adhered to, and grew and proliferated on the surface of the SIS framework. The bladder transitional epithelia covered the entire surface of the framework after nine days of culture in vitro, showing a single-layer cellular structure. The bladder transitional epithelia planted on the SIS framework formed a multi-layer structure after four weeks or eight weeks of transplant to the nude rats. The brown cellular plasma staining was observed, indicating a positive reaction to the immunohistochemical staining with cytokeratin.
Urothelial structures can be reconstructed in vitro and in vivo with tissue engineering technologies, which lays a technical foundation for further urinary tract reconstructing experiments.
利用组织工程技术开发一种重建尿路上皮结构的方法。
将幼犬的膀胱黏膜机械分离、切碎，并用胰蛋白酶消化。将获得的膀胱移行上皮用DKSFM培养，然后移植到小肠黏膜下层（SIS）支架表面。将上皮与小肠黏膜下层（SIS）材料的复合物种植于裸鼠背部皮肤下。在移植后第1、2、4和8周处死大鼠。对植入样本进行组织学和免疫组织化学检查。
膀胱移行上皮附着于SIS支架表面并生长增殖。体外培养9天后，膀胱移行上皮覆盖了支架的整个表面，呈现单层细胞结构。移植到裸鼠体内4周或8周后，种植在SIS支架上的膀胱移行上皮形成了多层结构。观察到棕色的细胞质染色，表明对细胞角蛋白免疫组织化学染色呈阳性反应。
利用组织工程技术可在体外和体内重建尿路上皮结构，为进一步的尿路重建实验奠定了技术基础。
