ENDOTHELIAL CELL OUTGROWTH FROM BLOOD

我们已经开发出新的方法，使我们能够从血液中获得内皮细胞的旺盛生长，从适量的外周血开始。这种生长具有健康的、静止的微血管内皮细胞的特征。我们建议这些将用于各种生物医学，生物医学工程和组织工程应用。为了验证这一点，我们将进行三项概念验证研究，以记录血液内皮生长的效用。为了说明使用这些细胞诊断内皮疾病的可行性，我们将在正常人、已知具有无功能突变血栓调节蛋白的人和血栓形成性素质的患者中检测它们的血栓调节蛋白活性。[2]为了说明使用它们来覆盖设备的可行性，我们将证明可以获得足够数量的内皮细胞，使血管移植物的管腔与内皮细胞层融合。为了说明它们在基因治疗中的效用，我们将证明我们可以表达修饰的凝血因子VIII（标记和无B结构域）的cDNA，然后进一步扩大转染/转导的内皮细胞群。

We have developed novel methods that allow us to reproducibly obtain an exuberant outgrowth of endothelial cells from blood, starting with reasonable quantities of peripheral blood. This outgrowth has the characteristics of healthy, quiescent microvascular endothelial cells. We propose that these will be of use in various biomedical, biomedical engineering, and tissue engineering applications. TO test this, we will conduct three proof-on-concept studies to document the utility of the endothelial outgrowth from blood. [1] To illustrate the feasibility of using these cells for diagnosis for endothelial-based disorders, we will assay them for thrombomodulin activity in normals, in persons known to have non-functional mutant thrombomodulin, and in patients with thrombotic diathesis. [2] To illustrate the feasibility of using them to coat devices, we will document that sufficient numbers can be obtained to line the lumen of vascular grafts with a confluent of layer of endothelial cells. [3] To illustrate their utility for gene therapy purposes, we will document that we can express cDNA for a modified coagulation factor VIII (marked and B domainless) and thereafter further expand the transfected/transduced endothelial cell population.