Angiogenesis Induction by Vasculogenic Progenitor Cells

描述(由申请人提供)：在冠状动脉和外周血管系统的缺血性疾病中诱导新的血管生长对于其治疗意义重大。体内血管化可以恢复细胞活力，诱导结构组织化，促进植入后的整合。胚胎干细胞在一种称为血管生成的过程中具有分化和形成新生血管的能力。我们已经展示了实验证据，证明存在来自人类胚胎干细胞(HES)的内皮细胞、平滑肌细胞和造血细胞(ESH)的共同祖细胞群体。使用一种新的方法，我们从HES株H9和H13(NIH代码分别为WA13和WA14)产生了一个增殖细胞群，显示出高水平的内皮细胞标记和内皮细胞和造血细胞的早期谱系标记上调。我们假设血管前体细胞可以在缺血组织中诱导血管生成。在这项研究中，我们将使用从我们实验室获得的HES系13、16和J3(NIH代码TE03、TE06和TE07)和H9和H13(NIH代码WA09和WA13)及其克隆。我们最初的重点将是鉴定和优化血管前体细胞在体外的稳定性，并进一步放大它们的生长和扩增，使它们适合细胞治疗的实施。第二个目标将是使用HES细胞系统研究人类血管生成。我们将利用延时视频记录，在体外研究和增强血管生成的自然机制，然后将血管生成基因转移到HES细胞。最后，我们将研究剪应力对HES细胞的影响，并将三维聚合物支架血管化。最后，我们计划在体内检测血管前体细胞。我们期望，这项研究将为使用HES细胞治疗世界上一些最主要的发病率和死亡率原因奠定有效的治疗方式的基础。

DESCRIPTION (provided by applicant): Induction of new vascular growth in ischemic disorders of the coronary and peripheral vasculature is significant for its therapeutic implications. Vascularization in vivo could restore cellular viability, induce structural organization and promote integration upon implantation. Embryonic stem (ES) cells have the capability to differentiate and form blood vessels de novo in a process called vasculogenesis. We have shown experimental evidence for the existence of a common progenitor cell population of endothelial, smooth muscle and hematopoietic (ESH) cells, derived from human embryonic stem (hES) cells. Using a novel approach, we generated from hES lines H9 and H13 (NIH code WA13 and WA14 respectively) a proliferating cell population, which exhibits high levels of endothelial markers and up-regulation of early lineage markers of both endothelial and hematopoietic cells. We hypothesize that vascular progenitor cells can induce angiogenesis in ischemic tissues. For this research we will use the hES lines derived in our laboratory 13, 16 and J3 (NIH code TE03, TE06 and TE07) and H9 and H13 (NIH code WA09 and WA13) and their clones. Our initial focus will be to characterize and optimize the stability of vascular progenitor cells in vitro, and further amplify their growth and expansion to make them suitable for cell therapy implementation. A second goal will be to study human vasculogenesis using the hES cell system. We will study and enhance the natural mechanism of angiogenesis, in vitro, using time-lapse video records, then transfer angiogenic genes to hES cells. Finally we will study the effects of shear stress on hES cells and vascularize three dimensional polymeric scaffolds. Finally, we plan to examine the vascular progenitor cells in vivo. We expect that this research will establish the foundation for an effective therapeutic modality using hES cells for some of the most leading causes of morbidity and mortality in the world.