Elucidating the Effects of Shear and Confinement on Endothelial Cell Differentiation

项目总结 虽然循环内皮细胞(ECs)显示出潜在的血管损伤部位，帮助其修复， 它们很难在体外分离和扩大，需要有潜在致病作用的动物产品。 它们被用作血管疗法。为此，包括人类在内的人多能干细胞(HPSC) 胚胎干细胞(HESCs)和人类诱导多能干细胞(HiPSCs)是一种潜在的更新来源 ECS。虽然它们是从hPSC衍生出来的，但用来诱导它们的化学线索 由于成熟培养液中使用的动物血清的内在变异性，分化是不明确的。在……里面 此外，与血管发育相关的物理线索的影响，如限制和剪应力 对于EC命运的决定和动/静脉的规范还有待阐明。这项提议的目标是 研究是以临床相关的方式实现动脉或静脉内皮细胞的同质性群体。我们的 目标是：(1)建立化学定义的差异化协议，以促进欧共体承诺 HPSCs；(2)通过在化学定义的培养条件下的限制来指导EC的命运，以及(3)确定其影响 切应力对EC动/静脉规格的影响。为了实现这些目标，高度跨学科的方法 包括干细胞和血管生物学领域以及工程学原理。成功 这些目标的完成将扩大我们对推动干细胞命运的物理化学因素的理解， 在将分化的细胞用作血管治疗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.