改良静态细胞种植方法在培育心血管组织工程材料中的价值

Tissue engineering of cardiovascular tissue using modified static ceeding method

目的评价改良静态细胞种植方法在培育二维心血管组织工程补片中的价值。方法将人脐带动脉壁平滑肌细胞以静态细胞种植方法种植在聚-4-羟基丁酸酯材料构建的补片支架材料上,制备组织工程补片。分别采用两种方法进行培育:静态培育为细胞材料复合物体外静态培育21 d;动态培育为细胞材料复合物先体外静态培育14 d,再置入生物反应器中继续动态组织培育7 d。对补片组织进行组织学、扫描电镜检查。免疫组化分析细胞外基质合成。结果静态细胞种植后经体外动态培育后,平滑肌细胞均匀地黏附于材料表面,细胞保持生长活力,并浸润生长入材料内部,形成均匀组织。结论改良静态细胞种植方法在二维组织工程支架材料上可达到细胞及组织的均匀分布。

Objective To evaluate the application of modified static cell seeding method in tissue engineering of 2- dimensional cardiovascular patch. Methods Patch scaffolds fabricated from poly- 4- hydroxy- butyrate（ P4HB） polymers were seeded with human umbilical artery myofibroblasts（ HUAMs） using static cell seeding method. In static cultivation group,the cell- polymer patch constructs were cultured in vitro for 21 days under static cell culture condition. In dynamic group,the cell- seeded patch constructs were incubated in vitro for 14 days under static culture conditions and were subsequently transferred into a bioreactor for another culture of 7 days. Morphological characterization of tissue engineered（ TE） patches was observed included histology and scanning electron microscopy（ SEM）. Extracellular matrix（ collagen） formation was analyzed by immunohistochemistry. Results After static cell seeding and subsequently conditioning in the bioreactor,the fibroblast cells were well adhered to the surface of P4HB polymer,the cells were mostly viable,had grown into the pores,and had formed uniform tissue on the patch constructs. Conclusion Using modified static cell seeding method,the uniform distribution of cells and tissue on the 2- dimensional tissue engineered structures can be achieved.