组织工程心脏瓣叶的体外培养

目的 探讨瓣叶支架种植细胞后,不同体外培养时间瓣叶的生成情况. 方法杂种猪9只,取主动脉,分离内皮细胞、成纤维细胞和平滑肌细胞,培养扩增.将成纤维细胞、平滑肌细胞和内皮细胞按顺序种植在预湿处理的支架材料上,以首次种植成纤维细胞、平滑肌细胞为起点,取7天、14天和28天样本使用戊二醛固定,用扫描电子显微镜(SEM)检测,对培养28天的样本进行组织学检查. 结果 SEM检测显示:随着培养时间的延长,细胞数量增加,细胞间连接出现,并有基质生成;至28天,瓣膜支架上可见大量细胞黏附,细胞融合成片,表面可见基质形成.支架的细胞覆盖率也随时间的延长明显增加.组织学检测见大量细胞黏附材料,细胞已长入材料中部. 结论体外培养的瓣叶存在生长活性,使用组织工程方法有可能生成组织工程心脏瓣膜.

In vitro experience with tissue engineered heart valve leaflets

Objective To investigate the possibility of creation of tissue engineered heart valve leaflets in vitro . Methods Aorta were obtained from 9 hybrid young pigs. The endothelial cell, fibroblast and smooth muscle cells were isolated and cultured to get enough cell. The expanded fibroblast, smooth muscle cell,and endothelial cells were seeded on the polymers sequentially. The cell polymer constructs were sent for scanning electron microscopy(SEM) examination after cultured for 7, 14, and 28 days. Histological examination were performed after the cell polymer constructs cultured for 28 days. Results SEM showed that the number of cells on the polymers increased as the culture time prolonged, with the formation of matrix. After 28 days, there were a great number of cells and large amount of matrix on the scaffolds. The confluent cell had covered a large area of the polymers. Hematoxylin and eosin(HE) stain showed large amount of cells attached to the polymers. Conclusion With the viability of the cultured cellular scaffolds,it is possible to create tissue engineered heart valve leaflets in vitro.