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重复获得大量内皮细胞的生长，从适量的外周血液开始。这种突起具有健康、静止的微血管内皮细胞的特征。我们认为，这些将在各种生物医学、生物医学工程和组织工程应用中使用。为了测试这一点，我们将进行三项概念验证研究，以证明血液中产生的内皮细胞的效用。[1]为了说明使用这些细胞诊断基于内皮的疾病的可行性，我们将在正常人、已知患有无功能突变的血栓调节蛋白的人和血栓形成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的数量来使血管移植物的管腔与内皮细胞层的汇合形成线。[3]为了说明它们用于基因治疗的有效性，我们将证明我们可以表达修饰的凝血因子VIII(标记的和无B结构域的)的cDNAs，然后进一步扩大转基因/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.