Elucidating the Effects of Shear and Confinement on Endothelial Cell Differentiation

阐明剪切和限制对内皮细胞分化的影响

• 批准号: 9195211
• 负责人: Quinton Smith
• 金额: $ 3.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9195211
• 关键词: Adherent Culture; Affect; Animals; Automobile Driving; Biology; Blood Vessels; Brachyury protein; Cardiovascular Diseases; Cause of Death; Cell Differentiation process; Cell Line; Cell Polarity; Cell Separation; Cells; Chemicals; Coupled; Coupling; Cues; Derivation procedure; Embryo; Embryonic Development; Endothelial Cells; Engineering; Environment; Epidemic; Event; F-Actin; Gene Expression; Generations; Goals; Home environment; Human; In Vitro; Injury; Kinetics; Lead; Left; Malignant Neoplasms; Mechanics; Methods; Microfluidics; Pathway interactions; Patients; Pattern; Perfusion; Pericytes; Phenotype; Phosphotransferases; Pluripotent Stem Cells; Population; Process; Protocols documentation; Research; Research Personnel; Role; Serum; Serum-Free Culture Media; Signal Transduction; Site; Sorting - Cell Movement; Source; Staging; Stimulus; Surface; System; Therapeutic; Tissues; United States; Vascular Diseases; Venous; cell fate specification; cell injury; cell type; clinically relevant; human embryonic stem cell; in vivo; induced pluripotent stem cell; interdisciplinary approach; novel; progenitor; protein expression; receptor; repaired; shear stress; stem cell differentiation; stem cell fate; stem cells; tool

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.

项目摘要 循环内皮细胞（ECS）显示出可能在其维修部位的病情部位的可能性时， 它们很难在体外分离和扩展，需要潜在的致病性动物产品 它们用作血管疗法。为此，人类多能干细胞（HPSC）包括人 胚胎（HESC）和人类诱导的多能干细胞（HIPSC），是潜在的更新来源 ECS。虽然已经实现了它们从HPSC衍生的，但化学线索用于诱导其 由于成熟培养基中使用的动物血清的固有变异性，分化的定义不明。在 补充，物理线索与发育中的脉管系统相关的效果，例如限制和剪切应力 尚未阐明朝着命运决策和动脉/静脉规范。这个提议的目标 研究是以临床相关的方式实现均质的动脉或静脉EC。我们的 目的是：（1）建立一种化学定义的分化方案，该方案促进EC的承诺 HPSC; （2）在化学定义的培养条件下通过限制来指导EC命运，（3）确定效果 对EC动脉/静脉规范的剪切应力。为了实现这些目标，一种高度跨学科的方法 采用，结合了干细胞和血管生物学领域以及工程原理。成功的 这些目标的完成将扩大我们对驱动干细胞命运的生理化学因素的理解， 将分化细胞用作血管疗法的影响很大。