In vitro functional testing of endothelial progenitor cells that overexpress thrombomodulin.
In vitro functional testing of endothelial progenitor cells that overexpress thrombomodulin.
复制标题
DOI：
10.1089/ten.tea.2010.0631
复制
发表时间：
2011-05
影响因子：
--
通讯作者：
J. Stroncek;Yujing Xue;N. Haque;J. Lawson;W. Reichert
中科院分区：
文献类型：
--
作者：
J. Stroncek;Yujing Xue;N. Haque;J. Lawson;W. Reichert
这项研究通过基因疗法过表达血栓调节蛋白(TM)来增强内皮祖细胞(EPC)的血栓抵抗力。血栓调节蛋白(TM)是一种具有强大抗凝特性的内皮细胞膜糖蛋白。从确诊的冠状动脉疾病患者的外周血中分离出晚期生长的内皮祖细胞，并将其与含有人TM的腺病毒载体转染。采用EPC转染条件以最大化TM的表达、转染率和细胞存活率。TM过表达的EPC比天然EPC和转染腺病毒控制载体表达β-半乳糖苷酶的EPC的活化蛋白C产生速率增加了五倍(p<；0.05)。与天然内皮祖细胞相比，TM上调导致血小板粘附性显著降低三倍，与I型胶原涂层孔相比，TM上调导致血小板粘附性降低12倍。此外，转TM基因的内皮祖细胞与全血孵育后的凝血时间比天然细胞显著延长19%(p<；0.05)。这些数据表明，TM过表达有可能改善患者来源的内皮细胞用于内皮化应用的抗血栓性能。
This study investigated the augmentation of endothelial progenitor cell (EPC) thromboresistance by using gene therapy to overexpress thrombomodulin (TM), an endothelial cell membrane glycoprotein that has potent anti-coagulant properties. Late outgrowth EPCs were isolated from peripheral blood of patients with documented coronary artery disease and transfected with an adenoviral vector containing human TM. EPC transfection conditions for maximizing TM expression, transfection efficiency, and cell viability were employed. TM-overexpressing EPCs had a fivefold increase in the rate of activated protein C production over native EPCs and EPCs transfected with an adenoviral control vector expressing β-galactosidase (p<0.05). TM upregulation caused a significant threefold reduction in platelet adhesion compared to native EPCs, and a 12-fold reduction compared to collagen I-coated wells. Additionally, the clotting time of TM-transfected EPCs incubated with whole blood was significantly extended by 19% over native cells (p<0.05). These data indicate that TM-overexpression has the potential to improve the antithrombotic performance of patient-derived EPCs for endothelialization applications.