小肠黏膜下层无细胞基质作为阴道平滑肌细胞载体的可行性  

Feasibility of small intestinal submucosa acellular matrix as vaginal smooth muscle cell carrier

作　　者：李庆林 刘伟[2] 李文芳[2] 刘德伍[2] 章杰[2] 

机构地区：[1]江西省皮肤病专科医院,江西省南昌市330006 [2]南昌大学第一附属医院,江西省南昌市330006

出　　处：《中国组织工程研究与临床康复》2010年第47期8741-8746,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金项目(30860294);江西省科技厅支撑项目~~

摘　　要：背景:目前国内外用于阴道组织工程研究的主要支架材料聚乙醇酸存在降解过快等缺陷。天然脱细胞支架材料尤其是小肠黏膜下层逐渐成为组织工程研究的重点。目的:探索用猪小肠黏膜下层基质作为组织工程学阴道细胞载体的可行性。方法:取新西兰雌兔,分离出阴道平滑肌组织块,组织块+酶消化法原代培养阴道平滑肌细胞。体外培养传代后作为种子细胞接种于自制猪小肠黏膜下层基质体外联合培养,倒置显微镜动态观察细胞形态及生长增殖情况,分别于1,2,3,4周时取标本,行组织学检查。结果与结论:①体外成功培养出阴道平滑肌细胞,倒置显微镜下,见培养的阴道平滑肌细胞呈现长梭状,细胞集结于培养皿上形成典型的"峰和谷"样构型。②黏膜下层无细胞基质外观呈白色,半透明,有一定韧性。苏木精-伊红染色未见细胞成分存在。③阴道平滑肌细胞-肠黏膜下层标本切片苏木精-伊红染色后,光镜下可见细胞成分逐渐增多,由表浅向深层部位生长。④阴道平滑肌细胞-肠黏膜下层标本切片采用抗兔平滑肌α-肌动蛋白单克隆抗体免疫组化染色后,均可见抗兔α-Actin的阳性细胞。结果初步证明了猪小肠黏膜下层基质可作为一种平滑肌细胞载体。BACKGROUND：The main scaffold for vaginal tissue engineering study,polyglycolic acid,has rapid degradation and other defects.Acellular natural scaffold material,in particular small intestinal submucosa,is becoming a hot spot of tissue engineering research.OBJECTIVE：To explore the possibility of using porcine small intestinal submucosa acellular matrix as a cell carrier in vaginal tissue engineering.METHODS：The vaginal smooth muscle tissues were separated from New Zealand female rabbits,and cultured by tissue explant ＋ collagen digestion methods in vitro,then passaged and incubated into self-made porcine small intestinal submucosa matrix as a seed cell.Morphology,growth and proliferation of the cells were monitored under the inverted microscope.The cell-matrix complex was harvested at 1,2,3,4 weeks for histological examination.RESULTS AND CONCLUSION：①The vaginal smooth muscle cells were successfully cultured in vitro.The cells were fusiform and aggregated in Petri dish,forming ＂peak-valley＂ structure.② The submucosa acellular matrix was white,semitransparent and tough.No cell component could be seen by hematoxylin-eosin staining.③ Specimens of vaginal smooth muscle cell-intestinal submucosa were stained by hematoxylin-eosin,cell components were increasing gradually and grew from surface to deep site under light microscope.④ Specimens of vaginal smooth muscle cell-intestinal submucosa were stained by monoclonal antibody of anti-rabbit smooth muscle a-actin by immunohistochemistry.The a-actin positive cells can be seen.Results preliminarily prove that porcine intestinal submucosa matrix can be used as smooth muscle cells carrier.

关 键 词：阴道平滑肌细胞 组织工程 猪小肠黏膜下层 细胞载体 支架材料 

分 类 号：R318[医药卫生—生物医学工程]