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目的 探索以液态胶原作为支架材料于体外构建组织工程化心肌组织。方法 将分离并培养的新生大鼠心肌细胞与添加了基质因子的液态Ⅰ型胶原混合 ,在环形槽中铸成环状心肌细胞 /胶原条带。构建的条带在环形槽中培养 7d后取出 ,在对其施加 10 %的静态拉伸的情况下 ,再培养 7d。通过光学显微镜、常规HE染色、免疫组织化学染色和透射电镜观察等对构建的组织工程化心肌组织进行评价。结果 构建的工程化心肌组织在施加静态拉伸的第 2天可观察到局部跳动的区域 ,随时间的延长跳动的区域逐渐增多 ,强度逐渐增大 ,并最终形成一致跳动。组织学和免疫组织化学染色研究结果显示 ,在构建的工程化心肌组织中 ,心肌细胞比较均匀地分布在整个条带中 ,且多数沿拉伸方向伸展 ,胞核呈长梭形 ,其形态类似于天然成熟心肌组织。透射电镜观察显示 ,条带中心肌细胞含有排列整齐的肌原纤维 ,并沿着细胞的长轴方向伸展 ,肌小节结构和Z带清晰可见。结论用液态Ⅰ型胶原可以构建类似天然心肌组织的工程化心肌组织。
Objective To explore the construction of tissue engineered myocardium with liquid collagen as scaffold material in vitro. Methods The neonatal rat cardiomyocytes isolated and cultured were mixed with liquid type I collagen supplemented with matrix factor and cast into circular cardiomyocytes / collagen bands in the annular groove. The constructed strip was removed after 7 days of culture in an annular groove and cultured for a further 7 days with 10% static stretching applied thereto. The constructed tissue engineered myocardium was evaluated by light microscopy, routine HE staining, immunohistochemistry and transmission electron microscopy. Results In the constructed myocardial tissue, local beating was observed on the second day after applying static stretching. The area of beating increased gradually with the extension of time, and the intensity gradually increased and eventually formed a consistent beat. Histological and immunohistochemical staining results showed that in the constructed engineered myocardium, cardiomyocytes were distributed more evenly in the whole band, most of them stretched along the stretching direction, and the nucleus was long fusiform and its morphology was similar In natural mature myocardial tissue. Transmission electron microscopy revealed that myofibroblasts in the strips contained neat myofibrils and extended along the long axis of the cells, and the structure of the myotubes and the Z band were clearly visible. Conclusions Engineered myocardial tissue resembling native myocardial tissue can be constructed using liquid type I collagen.