体外构建工程化心肌组织的实验研究  

Experimental study of engineered myocardial tissue in vitro

作　　者：李军杰[1] 吕安林[1] 刁繁荣[1] 苑媛[1] 张荣庆[1] 胡小菁[1] 

机构地区：[1]第四军医大学西京医院心内科,陕西西安710032

出　　处：《心脏杂志》2007年第4期421-423,427,共4页Chinese Heart Journal

摘　　要：目的探索以胶原海绵为支架,新生大鼠原代心肌细胞为种子细胞,于体外构建工程化心肌组织。方法用胰酶消化法分离新生大鼠心肌细胞,经初步纯化,然后接种于长方形片状胶原海绵中,将此细胞/胶原复合物置于细胞培养箱中,培养25 d,采用常规苏木精-伊红(HE)染色、免疫组织化学染色和透射电子显微镜等方法评价该工程化心肌组织片的形态和功能。结果构建的工程化心肌组织片先是出现局部自发收缩,随时间延长,自发收缩区域增多、强度增大,整个心肌片出现同步收缩,此收缩活动可持续25 d。检测结果显示,工程化心肌组织内细胞间连接广泛存在,细胞多呈纵向分布,胞核呈长圆形;多数细胞胞浆内α-肌节肌动蛋白阳性;细胞间连接清晰可辨,细胞内的肌原纤维排列整齐,可见到肌小节结构和Z线。结论用新生大鼠原代心肌细胞作为种子细胞、胶原海绵为支架,于体外可构建组织形态与生理功能类似于成熟大鼠心肌组织的工程化心肌组织。AIM To construct engineered myocardial tissue （EMT） using collagen sponge and neonatal rat primary cardiomyocytes. METHODS Neonatal rat cardiocytes were isolated using 2.5 g/L trypsin solution, purified, added to the collagen sponge segment placed in the rectanglar well made of glass and cultured in the incubator. Microscopy, routine HE staining, immunohistochemical staining and transmission electron microscopy were EMTs were seen not long after applied to assess the EMTs. RESULTS Spontaneous beating areas in the seeding. More beating areas could be seen afterwards with beats becoming stronger and stronger. Synchronously beating of the EMTs were finally observed. HE staining and immunohistochemical analyses showed that cardiocytes distributed equally within the 3-dimensional matrix, that intercellular contacts were very widespread and that the majority of the cardiocytes stretched in the EMTs. Immunohistochemical staining showed that α-actins in most cardiocytes were positive. Transmission electron microscopy showed that intercellular contacts were tight, that the cells in the EMTs contained well arranged myofibrils paralleled to the longitudinal cell axis and that clearly defined sarcomeres and Z lines were observed. CONCLUSION Using collagen sponge and neonatal rat cardiocytes EMTs can successfully be constructed in vitro, the histological and physiological properties of which resemble native cardiac tissue.

关 键 词：新生大鼠原代心肌细胞 胶原海绵 工程化心肌组织 

分 类 号：R540.27[医药卫生—心血管疾病]