Human blood vessel-derived endothelial progenitors for endothelialization of small diameter vascular prosthesis.
Human blood vessel-derived endothelial progenitors for endothelialization of small diameter vascular prosthesis.
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DOI：
10.1371/journal.pone.0007718
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发表时间：
2009-11-05
期刊：
影响因子：
3.7
通讯作者：
Hardikar AA
中科院分区：
文献类型：
--
作者：
Ranjan AK;Kumar U;Hardikar AA;Poddar P;Nair PD;Hardikar AA
急性血栓形成和内膜增生导致的冠状动脉旁路移植失败一直是涉及小直径人工血管的外科手术中的主要挑战。有人建议用患者自身的内皮细胞涂覆合成移植物，以提高搭桥手术的通畅率和总体成功率。我们从人隐静脉/乳动脉的剩余碎片中分离出内皮祖细胞（EPC）。我们证明 EPC 可以在低密度培养条件下扩增以产生数百万个细胞。暴露于高密度条件会诱导分化为内皮细胞表型。 EPC 衍生的内皮细胞表达 CD144high、CD31 和 vWF。然后，我们评估了分化内皮细胞在小直径膨胀聚四氟乙烯 (ePTFE) 管上粘附和生长的能力。由于 ePTFE 管具有高度疏水性，我们优化了方案，在 ePTFE 管的管腔表面引入亲水基团。我们在此演示了一个逐步方案，其中涉及引入亲水部分并用特定的 ECM 成分进行涂层，这些成分支持内皮细胞的粘附，但不支持血小板的粘附。我们的数据证实，从成人血管获得的内皮祖细胞可以在无异种蛋白的条件下在体外扩增，可用于小直径 ePTFE 移植物的内皮化。这些内皮化移植物可能代表了一种有前途的治疗策略，可改善心脏搭桥手术中小口径血管移植物的临床结果。
Coronary bypass graft failure as a result of acute thrombosis and intimal hyperplasia has been the major challenge in surgical procedures involving small-diameter vascular prosthesis. Coating synthetic grafts with patients' own endothelial cells has been suggested to improve the patency rate and overall success of bypass surgeries. We isolated endothelial progenitor cells (EPCs) from leftover pieces of human saphenous vein/mammary artery. We demonstrate that EPCs can be expanded to generate millions of cells under low-density culture conditions. Exposure to high-density conditions induces differentiation to endothelial cell phenotype. EPC–derived endothelial cells show expression of CD144high, CD31, and vWF. We then assessed the ability of differentiated endothelial cells to adhere and grow on small diameter expanded polytetrafluoroethylene (ePTFE) tubings. Since ePTFE tubings are highly hydrophobic, we optimized protocols to introduce hydrophilic groups on luminal surface of ePTFE tubings. We demonstrate here a stepwise protocol that involves introduction of hydrophilic moieties and coating with defined ECM components that support adhesion of endothelial cells, but not of blood platelets. Our data confirms that endothelial progenitors obtained from adult human blood vessels can be expanded in vitro under xenoprotein-free conditions, for potential use in endothelialization of small diameter ePTFE grafts. These endothelialized grafts may represent a promising treatment strategy for improving the clinical outcome of small-caliber vascular grafts in cardiac bypass surgeries.
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