脱细胞组织基质和自身细胞构建组织工程移植物  

Tissue-engineering graft constructed by autologous cells and heterogenous acellularized matrix

作　　者：黄惠民[1] 吴少峰[1] 任洪[1] 

机构地区：[1]上海交通大学医学院附属新华医院上海儿童医学中心胸心外科,200127

出　　处：《上海医学》2006年第5期315-318,F0003,共5页Shanghai Medical Journal

基　　金：上海市自然科学基金资助项目(99ZB14018)

摘　　要：目的采用脱细胞组织基质作为支架,自身内皮细胞和平滑肌细胞作为种子细胞,构建组织工程移植物。方法取小猪颈动脉,采用酶消化法分别得到内皮细胞和平滑肌细胞,于体外培养、扩增,获得足够量的种植细胞;另取羊主动脉,经多步骤脱细胞处理,获得羊脱细胞组织基质;将培养的猪内皮细胞和平滑肌细胞分别种植于羊脱细胞组织基质,体外培养3周后移植至细胞供体猪。结果培养、扩增的细胞经鉴定,内皮细胞和平滑肌细胞的特异性标志分别呈阳性反应;脱细胞组织基质中胶原纤维和弹性纤维保持原来的形态和结构,力学性能良好;细胞种植后移植至细胞供体猪,3个月后处死,见移植物内表面光滑,无血栓形成,无钙化,内膜层与周围组织连续分布,显微镜下见大量生长细胞,弹力纤维数量与正常血管壁相似。结论体外培养的自身细胞和异种动物脱细胞组织基质可作为构建组织工程移植物的良好材料。Objective Autologous endothelial and smooth muscle cells, and heterogenous acellularized matrix were chosen as the seeding cells and framework respectively to construct the tissue-engineering graft. Methods The seeding cells were obtained from a piglet. Common carotid artery was dissected 2 cm in length in order to get endothelial and smooth muscle cells by trypsin and collagenase digestion respectively. The isolated cells were further cultured and expanded using routine cell culture technique. The acellularized matrix was made by an adult sleep aortic artery. After multisteps management, all the above mentioned cells of the aorta were removed with preservation of elastic and collagen fibers. The endothelial cells and smooth muscle cells were seeded by multipoint injection into the acellularized matrix and cultured for another 3 weeks. Then the seeded graft was transplanted to the cell-donated piglet. Results The cultured cells from the piglet were characterized as the endothelial cell and the smooth muscle cell by the presence of vWF, CD31 and α-actin on the cell surfaces, respectively with immunohistochemical technique. In acellularized matrix from the sheep, all the cellular components were extracted, with the elastic and collagen fibers kept on their original morphology and structure. The maximal load of acellularized matrix lowerd by 20% but the maximal tension remained well. 3 months after transplantation, the piglet was killed and the graft removed for histopathology observation. The inner surface of the graft was smooth without adhered thrombus and calcification. Under microscopy, abundant growing cells could be seen, the amount of elastic fiber simulated that of the normal vessel wall. Conclusions Cultured autologous endothelial and smooth muscle cells and heterogenous acellularized matrix from the animal can be provided as the choice of tissue-engineering graft construction.

关 键 词：血管组织工程 自体细胞 脱细胞组织基质 移植 

分 类 号：R318.0[医药卫生—生物医学工程]