Elucidating the Effects of Shear and Confinement on Endothelial Cell Differentiation

项目摘要 虽然循环内皮细胞（EC）显示出归巢血管损伤部位的潜力，有助于其修复， 它们难以在体外分离和扩增，需要具有潜在致病性的动物产品， 它们作为血管治疗剂的用途。为此，包括人多能干细胞（hPSC）在内的人多能干细胞（hPSC）被用于治疗癌症。 胚胎干细胞（hESC）和人诱导多能干细胞（hiPSCS），作为一个潜在的更新来源， EC。虽然已经实现了它们从hPSC的衍生，但是用于诱导它们的化学线索仍然存在。 由于成熟培养基中使用的动物血清的固有可变性，分化不明确。在 此外，与发育中的脉管系统相关的物理线索的影响，例如限制和剪切应力， 对EC命运决定和动脉/静脉规格尚未阐明。这一提议的目的是 研究的目的是以临床相关的方式获得动脉或静脉EC的同质群体。我们 目的是：（1）建立一个化学定义的分化协议，促进EC承诺， hPSC;（2）通过限制在化学限定的培养条件下引导EC命运，以及（3）确定 EC动脉/静脉质量标准的剪切应力。为了实现这些目标，一个高度跨学科的方法， 是采取，结合领域的干细胞和血管生物学以及工程原理。成功 这些目标的完成将拓宽我们对驱动干细胞命运的理化因素的理解， 在使用分化的细胞作为血管治疗剂方面具有相当大的影响。

PROJECT SUMMARY While circulating endothelial cells (ECs) show the potential to home to sites of vascular injury, aiding in its repair, they are difficult to isolate and expand in vitro, requiring animal products that are potential pathogenic, curtailing their use as vascular therapeutics. To this end, human pluripotent stem cells (hPSCs) including human embryonic (hESCs) and human induced pluripotent stem cells (hiPSCS), serve as a potential renewal source of ECs. While their derivation from hPSCs have been achieved, the chemical cues used to induce their differentiation is ill-defined due to the inherent variability in animal serums used in the maturation media. In addition, the effect of physical cues relevant to the developing vasculature such as confinement and shear stress towards EC fate decision and arterial/venous specification have yet to be elucidated. The goal of this proposed research is to achieve a homogenous population of arterial or venous ECs in a clinically relevant manner. Our aims are to: (1) Establish a chemically-defined differentiation protocol that promotes EC commitment from hPSCs; (2) guide EC fate via confinement in chemically-defined culture conditions and (3) determine the effect of shear stress on EC arterial/venous specification. To achieve these aims, a highly interdisciplinary approach is taken, incorporating the fields of stem cell and vascular biology as well as engineering principles. Successful completion of these aims will broaden our understanding of the physio-chemical factors driving stem cell fate, with considerable impact in using the differentiated cells as vascular therapeutics.