去细胞同种心脏瓣膜的再内皮细胞化  

Recellularization of decellularized autologous heart valves

作　　者：崔彬[1] 刘迎龙[1] 于存涛[1] 谢宁[1] 宋来凤[2] 

机构地区：[1]中国医学科学院中国协和医科大学,阜外心血管病医院心血管外科,北京市100037 [2]中国医学科学院中国协和医科大学,阜外心血管病医院病理科,北京市100037

出　　处：《中国组织工程研究与临床康复》2007年第23期4580-4582,共3页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:初步构建受体内皮细胞化的组织工程同种心脏瓣膜,减轻移植后免疫排斥反应,并探讨内皮细胞适宜的种植方法,增强内皮细胞与细胞外基质的黏附性。方法:实验于2004-06在阜外医院心血管外科先心病研究室完成。①实验材料:取冠状动脉搭桥患者术中剩余的大隐静脉(该材料是术中多余弃掉部分,患者知情同意),研究方案获伦理委员会批准。②实验方法:采用低渗液-去污剂(10g/L去氧胆酸)-核酸酶法去除同种心脏瓣膜组织中的全部细胞成分,构建去细胞的同种心脏瓣膜;取培养3～4代的成人大隐静脉内皮细胞,依照内皮细胞密度分为生理密度种植组:以1.5×108L-1密度种植;超生理密度种植组:以(4～5)×108L-1密度种植;生理密度反复种植组:以1.5×108L-1密度,反复种植3d。③实验评估:种植于脱去全部细胞的同种心脏瓣膜上,静态培养5d,扫描电镜观察内皮细胞的覆盖情况。结果:去细胞同种心脏瓣膜的细胞外基质有着良好的细胞黏附性;3种不同的内皮细胞种植密度,其中以内皮细胞超密度(4～5)×108L-1种植,静态培养5d,同种组织瓣膜表面基本覆盖单层内皮细胞,细胞间相互延伸汇合,形态结构良好。以生理密度种植内皮细胞覆盖率较低,细胞大多孤立存在,细胞间较少相互延伸汇合。生理密度反复种植3d,内皮细胞覆盖率低于超生理密度种植组,细胞间可见相互延伸汇合,细胞形态结构良好。结论:去细胞同种心脏瓣膜的再内皮细胞化保留了同种心脏瓣膜的正常结构、功能及良好的组织相容性,有望延长瓣膜的使用寿命,取得更好的临床疗效。AIM： To develop the tissue-engineered heart valves based on human decellular homograft matrix reseeded with autologous endothelial cells to lessen the immunological rejection after transplantation and investigate the ideal method to enhance the adhesion of endothelial cells and matrix. METHODS： The experiment was conducted in the Laboratory of Congenital Heart Disease, Fuwai Hospital for Cardiovascular Disease in June 2004. ①The great saphenous vein abandoned after the coronary bypass was selected after the informed consent of patients, and the study scheme was permitted by the Hospital Ethics Committee. ②For decellularization, the homologous heart valves were treated with the hypotension buffer degreaser （10 g/L deoxycholic acid）-nuclease to construct the decellular heart valves; the third and fourth generation saphenous endothelial cells were divided into three groups according to the density of cells： 1.5×10^8 cells/L group, 4-5×10^8 cells/L group and 1.5×10^8 cells/L group, in which the cells were cultivated for 3 days. ③The cells were seeded onto the decelluladzed human valves and cultivated for 5 days statically. Cell coverage was examined by scanning electron microscope. RESULTS： The extracellular matrix of decellularized human valves had good cell adhesion; After recellularization, the decellularized valves were almost covered by the endothelial cells with the density of 4-5×10^8 cells/L, with excellent morphology. The coverage rate of 1.5×10^8 cells/L group was low, and cells were separated, with little fusion. After three days cultivation, the condition was improved, but the coverage rate was still lower than that of 4-5×10^8 cells/L group, CONCLUSION： The recellularization of the decellular homograft valve reserves the normal structure, function of heart valve and good histocompatibility, It is hopeful to prolong the service life of valves and gain better clinical effect.

关 键 词：组织工程 同种心脏瓣膜 内皮细胞化 组织构建 

分 类 号：R318[医药卫生—生物医学工程]