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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。

基本信息

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

项目摘要

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，初级神经丛和随后由大小血管组成的血管树形成的机制很大程度上是未知的。小鼠 ES 细胞的体外分化系统可将复杂的血管发育过程分解为可实时监测的细胞事件。我们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由源自 ES 细胞的单个中胚层前体细胞分化而来。该培养系统鉴定了中胚层细胞群中的三能心血管祖细胞。在基质细胞层上产生的ES细胞衍生的内皮细胞集落已被用作体外模型来检查单个内皮细胞响应各种血管生成刺激（例如VEGF）的行为。 VEGF 信号显示可诱导内皮细胞的伸长和分散。我们鉴定了叉头型转录因子 Foxo1 作为内皮细胞伸长反应的调节分子，这可能是 Foxo1 缺陷小鼠胚胎中血管生成严重异常的原因。我们在 VE-cadherin 基因启动子/增强子的调控下产生了表达 VE-cadherin-VENUS 或紧密蛋白 5-VENUS 的 ES 细胞克隆。对 ES 细胞衍生的内皮细胞集落的延时分析揭示了内皮细胞的活跃运动。尽管内皮细胞具有高运动性，但基于 VE-钙粘蛋白的粘附连接和基于密蛋白 5 的紧密连接蛋白的完整性得以维持。在移动内皮细胞的前缘发现了细胞连接的动态重塑过程。内皮细胞运动的调节可能是细胞水平上驱动血管发育的重要过程。利用VE-cadherin启动子-Cre转基因小鼠进行的命运追踪分析表明，心脏缺血会激活先前存在的血管细胞和参与新生血管形成的骨髓细胞中的VE-cadherin启动子。我们还证明转录因子 c-Myb 的过度表达会增加内皮细胞群中造血前体细胞的频率，这为了解造血内皮细胞在小鼠胚胎发生过程中如何分化为确定的造血细胞谱系提供了线索。较少的