Differentiating Embryonic Stem Cells Toward Arterial and Venous Endothelial Cells

描述（由申请人提供）：胚胎干细胞（ESC）衍生的内皮细胞（EC）具有巨大的潜力，可用于各种治疗领域，如血管移植物的组织工程和缺血组织的血管再形成。还非常期望获得用于特定应用的功能性动脉或静脉EC的均质培养物。迄今为止，已经开发了各种方案来将ESC分化为血管EC。然而，使用目前的方法从ESC衍生的EC主要显示静脉表型。因此，迫切需要开发精细的动静脉分化方法来解决这一差距。基于血管发育的发现，我们假设胚胎干细胞来源的Flk 1 + Nrp 1+细胞作为动脉EC祖细胞。我们认为，这一亚群细胞群倾向于动脉分化，并可以选择指导动脉分化与环境因素相结合。我们的初步数据支持这一假设。本研究的目的是使用人类ESC进一步验证这一假设。然后，我们将设计最佳的体外环境，引导胚胎干细胞进入动脉和静脉细胞的命运，并比较它们在组织工程应用中的功能后果。具体而言，我们将：（1）验证Nrp 1可用于从干细胞中识别动脉EC祖细胞，并定义引导ESC进入动脉和静脉细胞命运的最佳体外环境。(2)分析体外和体内ESC源性动脉和静脉EC在组织工程构建物中形成相互连接的功能性血管网络的能力。(3)在体外和体内确定ESC衍生的动脉和静脉EC在组织工程血管移植物重塑中的功能后果。

DESCRIPTION (provided by applicant): Embryonic stem cells (ESCs) derived endothelial cells (ECs) have enormous potential to be used in a variety of therapeutic areas such as tissue engineering of vascular grafts and re-vascularization of ischemic tissues. It is also much desired to obtain homogeneous culture of functional arterial or venous ECs for specific applications. To date, various protocols have been developed to differentiate ESCs toward vascular ECs. However, ECs derived from ESCs using current methods display predominantly venous phenotype. Therefore, developing refined method of arterial-venous differentiation is critically needed to address this gap. Based on the findings of vascular development, we hypothesize that embryonic stem cell derived Flk1+Nrp1+ cells serve as arterial EC progenitors. We think that this subset cell population is predisposed to arterial differentiation and can be selected to guide arterial differentiation in combination with environmental cues. Our preliminary data support this hypothesis. The goal of this study is to further test this hypothesis using human ESCs. We will then engineer optimal in vitro environments that guide ESCs into arterial and venous cell fate and compare their functional consequences in tissue engineering applications. Specifically, we will: (1) Validate that Nrp1 can be used to identify arterial EC progenitor from stem cells and define optimal in vitro environments that guide ESCs into arterial and venous cell fate. (2) Analyze the ability of ESC-derived arterial and venous ECs to form interconnected functional vascular network in tissue-engineered construct both in vitro and in vivo. (3) Determine the functional consequences of ESC-derived arterial and venous ECs in the remodeling of tissue engineered vascular graft both in vitro and in vivo.