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

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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.