Induction of cardio-vascular progenitor cells and blood vessel formation by ES cell differentiation.

通过 ES 细胞分化诱导心血管祖细胞和血管形成。

• 批准号: 16390227
• 负责人: OGAWA Minetaro
• 金额: $ 8.77万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390227/
• 关键词: Vascular endothelial cells; Cardiomyocytes; Vascular smooth muscle cells; Mesoderm; Embryonic stem cells; Cell movement; Cell adhesion; Transcription factors; 試験管内分化; タイムラプス解析

The mechanism by which primary plexus and subsequent vascular tree which consists of large and small vessels form in the process of vertebrate embryogenesis is largely unknown. In vitro differentiation system of murine ES cells serves as a means to break down the complicated processes of vascular development into cellular events which can be monitored in real time. We have developed a culture system in which endothelial cells, cardiomyocytes and smooth muscle cells differentiate from a single mesodermal precursor cell derived from ES cells. This culture system identified tri-potent cardio-vascular progenitor cells in the mesodermal cell population.ES cell-derived endothelial cell colonies generated on stromal cell layer have been used as an in vitro model to examine the behavior of individual endothelial cells in response to various angiogenic stimuli such as VEGF. The VEGF signal was shown to induce elongation and dispersion of endothelial cells. We identified a forkhead-type transcri … More ption factor Foxo1 as a regulatory molecule in the elongation reaction of endothelial cells, which might account for the severe abnormality of angiogenesis found in foxo1-deficient mouse embryos.We produced ES cell clones which express VE-cadherin-VENUS or claudin 5-VENUS under the regulation of VE-cadherin gene promoter/enhancer. Time-lapse analysis of ES cell-derived endothelial cell colonies revealed an active movement of endothelial cells. Integrity of the VE-cadherin-based adherens junction and the claudin 5-based tight junctin was maintained despite the high motility of endothelial cells. A dynamic remodeling process of cell junctions was identified at the leading edges of moving endothelial cells. Modulation of endothelial cell motility might be an important process which drives the vascular development at the cell level.Fate tracing analyses by using VE-cadherin promoter-Cre transgenic mice showed that cardiac ischemia activate VE-cadherin promoter in both pre-existing vascular cells and bone marrow cells involved in neovascularization. We also demonstrated that over-expression of a transcription factor c-Myb increases the frequency of hemogenic precursors in the endothelial cell population, which provides a clue to how hemogenic endothelial cells commit to definitive hematopoietic cell lineages during the mouse embryogenesis. Less

在脊椎动物胚胎发生过程中，原发丛和随后的血管树由大小vissel形成的机制在很大程度上是未知的。鼠ES细胞的体外分化系统是将血管发育的复杂过程分解成细胞事件的一种手段，可以实时监测。我们已经开发了一种培养系统，其中内皮细胞，心肌细胞和平滑肌细胞与来自ES细胞的单一中胚层前体细胞区分开。该培养系统确定了中胚层细胞群中的三动力心血管祖细胞。ES细胞衍生的内皮细胞菌落在基质细胞层上产生的内皮细胞菌落已被用作体外模型，以检查单个内皮细胞在响应各种血管疾病刺激的响应中的单个内皮细胞的行为。 VEGF信号显示出诱导内皮细胞的伸长和分散。 We identified a forkhead-type transcri … More ption factor Foxo1 as a regulatory molecule in the elongation reaction of endothelial cells, which might account for the severe abnormality of angiogenesis found in foxo1-deficiency mouse embryos.We produced ES cell clones which express VE-cadherin-VENUS or claudin 5-VENUS under the regulation of VE-cadherin gene promoter/enhancer.对ES细胞衍生的内皮细胞菌落的延时分析显示，内皮细胞的活动运动。基于VE-钙粘着蛋白的粘附连接和基于Claudin 5的紧密联合蛋白的完整性保持了内皮细胞的高运动。在移动内皮细胞的领先边缘确定了细胞连接的动态重塑过程。内皮细胞运动的调节可能是一个重要的过程，它可以在细胞水平上驱动血管发育。使用VE-钙粘蛋白启动子-CRE-CRE-CRE-CRE转基因小鼠进行进食，表明在涉及涉及的血管片细胞和骨髓化细胞中，心脏缺血激活了VE-Cadherin启动子。我们还证明，转录因子C-MYB的过表达增加了内皮细胞群中血液生成前体的频率，这为小鼠胚胎发生过程中血肿内皮细胞如何致力于造血性造血细胞谱系提供了线索。