人脐静脉内皮细胞用于构建组织工程心脏瓣膜的实验研究

Construction of tissue-engineered heart valve by seeding human umbilical vein endothelial cells on decellular porcine aortic valve

目的:以人脐静脉血管内皮细胞(HUVECs)为种子细胞,体外构建组织工程心脏瓣膜(TEHV),并对其分泌纤溶物质——组织型纤溶酶原激活物(t-PA)及其抑制物(PAI-1)的作用进行观察。方法:获取并扩增HUVECs,种植在猪脱细胞主动脉瓣叶上,体外静态构建TEHV。观察内皮细胞的形态学变化和生长状况。收集瓣膜培养液,采用发色底物法检测瓣膜内皮细胞分泌的t-PA和PAI-1活性。结果:以猪脱细胞主动脉瓣叶作支架,HUVECs做种子细胞,体外成功构建TEHV,种植的HUVECs在瓣膜表面长成一层连续的细胞层,生长状态良好。瓣膜表面的内皮细胞能够分泌纤溶物质t-PA和PAI-1。结论:HUVECs种植在猪脱细胞瓣膜支架上可以构建TEHV,且瓣膜内皮细胞能够分泌t-PA和PAI-1。

Objective:To develop the tissue-engineered heart valve(TEHV) by seeding human umbilical vein endothelial cells(HUVECs) on decellular porcine aortic valve and to investigate its anti-coagulation function. Methods:The porcine aortic valve was decellularized using Trypsin.EDTA.Triton X-100.RNase and DNase treatment. HUVECs were cultured and seeded on decelluar valve to create TEHV in vitro. Histology and growth condition of cells were observed. Contents of tissue plas-minogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) of endothelial cells were determined with the methods of colorsubstrate. Results:In vitro TEHV by seeding HUVECs on decellular porcine aortic valve was created successfully. Endothelial cells formed a continuous monolayer on the surface of valve and the cells can secrete t-PA and PAI-1. ConcuIsion:TEHV can be created using HUVECs as seed cells seeded on decellular porcine aortic valve, and the endothelial cells can secrete t-PA and PAI-1.