膀胱粘膜下层无细胞基质作为细胞载体的可行性研究  

A study of vesical submucosa acelluler matrix as cell deliverer

作　　者：耿红全[1] 唐达星[2] 陈方[1] 周昕[1] 吴湘如[3] 谢华[1] 徐卯升[1] 刘国华[1] 

机构地区：[1]上海第二医科大学附属新华医院上海儿童医学中心小儿外科,200092 [2]浙江大学医学院附属浙江儿童医院外科 [3]上海第二医科大学附属新华医院上海儿童医学中心,病理科200092

出　　处：《中华小儿外科杂志》2003年第5期421-423,F003,共4页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金(301709361);上海市"曙光计划";上海市教委重大科研基金(99ZD06)资助

摘　　要：目的 探讨膀胱粘膜下层无细胞基质作为组织工程学细胞载体的可行性。方法 采用胶原酶自膀胱组织分离出平滑肌细胞,在DMEM(含10％胎牛血清)中培养、传代。自膀胱显微解剖出膀胱粘膜下层,以0.5％SDS、双蒸水彻底洗涤后形成无细胞基质。平滑肌细胞以5.0×10~6/cm^2密度种植于膀胱无细胞基质,分别于1周、2周、3周、4周时取标本,HE染色、扫描电镜等观察细胞生长情况。结果 平滑肌细胞可在膀胱粘膜下层无细胞基质上粘附。1周时,平滑肌细胞主要位于膀胱粘膜下层无细胞基质的表面。第2、3周,细胞数量明显增多,并可见细胞向膀胱粘膜下层无细胞基质的中间部位侵入。第4周细胞数量减少。结论 膀胱粘膜下层无细胞基质可以作为细胞载体应用于组织工程学研究。该细胞-载体复合物的回植时间应在3周前。Objective To explore the possibility of using vesical submucosa acelluler matrix as cell deliverer in tissue engineering. Methods Smooth muscle cells were isolated from bladder tissues by collage- nase digestion, and than cultured and passaged in DMEM supplemented with 10% fetal bovine serum. Vesical submucosa was extracted by microdissection and was thoroughly washed in 0.5% SDS and dH2O. Smooth muscle cells were seeded onto the matrix at a density of 5.0×10~6 cells per cm^2. The cell-matrix complex was harvested at 1, 2, 3 and 4 weeks. The growth of SMCs was determined by HE stain and elec- tronic microscopic scan. Results The SMCs can adhere to bladder submucosa acelluler matrix and be locat- ed in the limited surface areas of matrix or just below the surface 1 week later. Quantity of the cells in- creased markedly and cells penetrated the inner site of the matrix after 2 and 3 weeks of cell seeding. But 4 weeks after seeding, the quantity of cells in the cell - matrix complex decreased. Conclusions Vesical sub- mucosa acelluler matrix can be used as cell deliverer in tissue engineering research. The cell-deliverer com- plex should be implanted to host within 3 weeks after cell seeding.

关 键 词：膀胱粘膜下层无细胞基质 细胞载体 可行性研究 胶原酶 细胞培养 组织工程 生物材料 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]