Differentiating Embryonic Stem Cells Toward Arterial and Venous Endothelial Cells

描述（由申请人提供）：胚胎干细胞（ESCs）衍生的内皮细胞（ECs）具有巨大的潜力，可用于各种治疗领域，如血管移植的组织工程和缺血组织的再血管化。对于特定的应用，也非常希望获得功能性动脉或静脉内皮细胞的均匀培养。迄今为止，已经制定了各种方案来区分ESCs和血管ECs。然而，使用当前方法从ESCs衍生的ECs主要显示静脉表型。因此，迫切需要发展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衍生的Flk1+Nrp1+细胞作为动脉EC祖细胞。我们认为，这一亚群细胞群易发生动脉分化，可以结合环境因素来指导动脉分化。我们的初步数据支持这一假设。本研究的目的是利用人类ESCs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ESCs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Nrp1可用于从干细胞中识别动脉内皮细胞祖细胞，并确定引导内皮细胞进入动脉和静脉细胞命运的最佳体外环境。(2)分析体外和体内组织工程构建中esc来源的动脉和静脉内皮细胞形成相互连接的功能血管网络的能力。(3)在体外和体内确定esc来源的动脉和静脉内皮细胞在组织工程血管移植物重塑中的功能影响。

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.