To establish a mucosal cell culture method, To make a cavity with a mucosal lining

在鼻、口、食道以及生殖和泌尿器官中，使用黏膜衬里的复杂组织进行重建是必要的。黏膜衬里组织在膀胱癌切除后的重建中尤为重要。今年的研究目标是：1)建立一种高效的兔舌下粘膜细胞培养方法；2)将培养的黏膜细胞注入肌腔内，然后附着于肌壁，并确认是否形成了由肌肉包裹的黏膜衬里空腔(类似于膀胱)；以及3)评估游离移植后是否获得了膀胱功能。细胞计数在培养第28天时平均为5~6×10~6个细胞。这是一个缓慢的生长速度，但本研究涉及到体内和体外的组织工程，所以培养的粘膜细胞回到体内后的增殖也是可以预期的。因此，我们得出结论，细胞数量是充足的。使用下面描述的方法将培养的粘膜细胞注入组织中。股直肌内置入6.5mL硅胶组织扩张器。在组织扩张器旁边插入用于注射培养细胞的硅胶管。在术后第7天注射细胞前，将细胞与纤维蛋白胶混合，将培养的细胞附着在肌肉上。从第4天开始用组织扩张器扩张。2周后，对细胞层进行组织学检查。在所有被检查的样品中都可以看到“年糕”的粘连模式。我们目前正在研究一种不使用纤维蛋白胶附着粘膜细胞的方法。

Reconstruction using mucosa-lined complex tissues is necessary in the nose, mouth, and esophagus, as well as the reproductive and urinary organs. Mucosa-lined tissue is particularly important in reconstruction of the bladder after excision cancer.The objectives of the studies for this year are as follows : 1)to establish a highly effective culture method using rabbit sublingual mucosal cells ; 2)to inject cultured mucosal cells into a cavity in muscle, which then become attached to the muscle wall, and to confirm whether a cavity with a mucosal lining that is enclosed by muscle (resembling the bladder) is formed ; and 3)to assess whether bladder function is acquired after free grafting.We established a mucosal cell culture method during 2003-2004. With regard to the cell count, a mean of about 5〜6x10^6 cells was obtained on Day 28 after the start of the culture. This is a slow growth rate, but this study is concerned with in vivo and ex-vivo tissue engineering, so proliferation of cultured mucosal cells after return to the body is also expected. Therefore, we concluded that the cell numbers were adequate.Injection of cultured mucosal cells into tissues was performed using the method described below. A 6.5 mL silicone tissue dilator was inserted into the rectus muscle of the thigh. A silicone tube for injecting cultured cells was inserted alongside the tissue dilator. Before cells were injected on postoperative Day 7, they were mixed with fibrin glue to attach the cultured cells to the muscle. Dilation with the tissue dilator was started from Day 4. After 2 weeks, the cell layer was examined histologically. A "rice cake" pattern of adhesion was seen in all the examined samples. We are currently investigating a method of attaching the mucosal cells that does not use fibrin glue.