Human Embryonic Stem Cell-Derived Endothelial Cells

呼吸道的运输需要胚胎和胎儿快速的血管形成和生长。 排泄物进入或排出胎儿的气体、营养物质和代谢废物，以支持快速成长的胎儿然而， 人类胚胎早期血管生成和血管生成的机制仍不清楚，原因是 显而易见的道德原因。最新研究证明血管内皮细胞在体外成功分化 来自小鼠和恒河猴的胚胎干细胞为研究提供了重要的工具 这样的机制。尽管如此，关于人类胚胎干细胞向内皮细胞分化的报道有限，而且 成功建立稳定的人ES细胞来源的内皮细胞系 目前还没有活动或血管扩张剂的报道。更重要的是，控制内皮细胞的机制 不同物种间胚胎干细胞的分化可能不同。因此，我们建议建立人胚胎干细胞来源的 具有血管生成能力的血管内皮细胞系(HESCDE)。四个具体的 AIMS将使用特征良好的人类ES细胞系(NIH人类ES细胞注册代码： WA01和WA09)：目的1.确定和优化诱导血管内皮细胞分化的条件 人ES细胞来源的类胚体(EBS)和人ES细胞的体外培养 目的：确定HESCDE细胞在体外是否有血管生成 碱性成纤维细胞生长因子(BFGF)和血管扩张剂一氧化氮 血管内皮生长因子(VEGF)的体外培养；目的3.确定HESCDE细胞是否具有 能够形成血管并在移植后整合到预先存在的血管中 严重联合免疫缺陷(SCID)米色小鼠；和目的4.比较 人脐静脉内皮细胞(HUVEC)、人胎盘动脉内皮细胞(HPAE)、 和人胎盘微血管内皮细胞(HPME)的基因芯片分析。 此外，为了进一步描述和定义HESCDE细胞的功能，我们将提供以下内容 项目I、II和IV的HESCDE单元与UAEC在静态状态下的行为比较(项目I)，下 动态剪应力(项目II)条件，并用作滋养层细胞分化的效应细胞(项目II) 四)。因此，这些研究将收集有关调控机制的潜在重要信息 人类早期胚胎发育过程中血管的内皮分化和形成。 此外，它还可能提供高纯度和无限的内皮细胞供应，不仅用于研究 调节血管生成的机制，但也用于临床应用，如组织移植 作为一种更持久和更复杂的基因疗法的载体。

Rapid vascular formation and growth in embryos and fetuses are required for transportation of respiratory gases, nutrients, and metabolic wastes into or out of fetuses to support the rapidly growing fetuses. However, the mechanisms underlying early vasculogenesis and angiogenesis in the human embryo remain poorly defined due to obvious ethical reasons. Recent studies demonstrating the successful differentiation of endothelial cells in vitro from mouse and rhesus monkey embryonic stem (ES) cells has provided an important tool for investigating such mechanisms. Nonetheless, reports on endothelial differentiation from human ES cells are limited, and the successful establishment of stable human ES cell-derived endothelial cell lines with potent angiogenic activities or vasodilator has not been reported. More importantly, the mechanisms controlling endothelial differentiation from ES cells may differ among species. Thus, we propose to establish human ES cellderived endothelial (HESCDE) cell lines with angiogenic capacity in vitro and in vivo. Four Specific Aims will be addressed using well-characterized human ES cell lines (NIH Human ES Cell Registry Code: WA01 and WA09): AIM 1. To determine and optimize conditions for inducing endothelial differentiation from human ES cells in vitro by treating human ES cell-derived embryoid bodies (EBs) and human ES cells with a defined differentiation medium; AIM 2. To determine whether these HESCDE cells have in vitro angiogenic capacities and produce nitric oxide, a vasodilator, in response to basic Fibroblast Growth Factor (bFGF) and Vascular Endothelial Growth Factor (VEGF) in vitro; AIM 3. To determine whether these HESCDE cells are capable of forming blood vessels and integrate into pre-existing blood vessels after transplantation into severe combined immunodeficient (SCID) beige mice; and AIM 4. To compare global gene expression in HESCDE cells with human umbilical vein endothelial, (HUVEC), human placental artery endothelial (HPAE), and human placental microvascular endothelial (HPME) cells using human cDNA microarray analysis. Additionally, to further characterize and define the functions of HESCDE cells, we will provide these HESCDE cells for Projects I, II & IV to compare their behaviors with UAEC in static (Project I), under dynamic shear stress (Project II) conditions, and used as effector cells for trophoblast differentiation (Project IV). Thus, these studies will glean potentially important information regarding the mechanisms that regulate endothelial differentiation and formation of blood vessels during early human embryonic development. Moreover, it will potentially provide highly purified and unlimited endothelial cell supply not only for studying mechanisms that regulate angiogenesis but also for clinical applications such as tissue transplantation therapies and as a vehicle for more enduring and sophisticated gene therapies.