Optimization of the Chemical-Physical Environments for Stem Cell Differentiation

干细胞分化的化学物理环境的优化

• 批准号: 7976461
• 负责人: SHU CHIEN
• 金额: $ 18.57万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7976461
• 关键词: Advanced Development; Blood Cells; Blood Vessels; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cell Count; Cell Culture Techniques; Cell Differentiation process; Cell Therapy; Cells; Chemicals; Clinical; Collaborations; Commit; Cues; Development; Devices; Differentiation and Growth; Elements; Environment; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblast Growth Factor; Foundations; Goals; Growth Factor; Healthcare; Heart; Heart failure; Hematological Disease; Hydrogels; In Vitro; Investigation; Knowledge; Laboratories; Life; Lung; Mechanics; Methods; Morphogenesis; Muscle Rigidity; Patients; Physical environment; Play; Process; Production; Property; Protein Array; Protein Microchips; Proteins; Protocols documentation; Regenerative Medicine; Research; Role; Screening procedure; Smooth Muscle Myocytes; Staging; Stem cells; Stretching; System; Technology; Testing; Tissue Engineering; Tissues; Translations; WA01 cell line; WA09 Cell Line; angiogenesis; cell growth; cell type; combinatorial; design; experience; human embryonic stem cell; improved; novel; public health relevance; scaffold; scale up; shear stress; stem cell differentiation; stem cell division; stem cell fate; tool; vasculogenesis

DESCRIPTION (provided by applicant): Pluripotent human embryonic stem cells (hESCs) can be differentiated in vitro into multiple cell types, including cardiovascular cells (cardiomyocytes, vascular endothelial and smooth muscle cells). The ability to control the growth and differentiation of stem cells in vitro is essential for the successful application of differentiated cells for cell-based therapies. The appropriate cell types committed to a desired lineage, together with the relevant elements of the cardiovascular system, can be used to treat cardiovascular diseases. The proposed research on the identification of the optimal condition for controlling hESC fate uses a novel medium-high throughput microarray platform composed of comprehensive chemical-physical microenvironments. These include (1) immobilized growth factors (GFs) and extracellular matrix proteins (ECMPs), (2) mechanical properties of the ECM, and (3) external mechanical forces acting on the cells. Current knowledge indicates that each of these parameters (i.e., GFs, ECMPs, substrate rigidities, as well as mechanical loadings) plays roles in regulating stem cell fate, but the efficiency is low when acting alone. Since stem cells experience the influence of multiple microenvironmental factors which change during the developmental stages, it is essential to examine the combinatory effects of multi-factorial complexities of the niches, as proposed in the current research. In the proposed research, we will investigate the combinatorial effects of ECM proteins (ECMPs) and GFs on the differentiation of hESCs (Federally approved WA01 and WA09 cell lines). We have designed a hydrogel system to control the rigidities of matrices, covering a range encountered in different tissues, for our ECMP array in stem cell culture. We will also incorporate the flow and stretch devices developed in our lab into the microarray system to assess the roles of external mechanical forces in regulating the cell fate of hESCs on the ECMP/GF/Rigidity platform. This novel combinatory microarray system allows the comprehensive testing of the chemical-physical microenvironment for the choice of optimal conditions for hESC growth and differentiation. The use of such optimally chosen hESCs for translation to cardiovascular tissue engineering will provide the opportunity to significantly advance the development of artificial vessels, angiogenesis patches, as well as cell replacement for heart failure, which will in turn improve the healthcare of patients with cardiovascular diseases and the wellbeing of our citizens. PUBLIC HEALTH RELEVANCE: Pluripotent human embryonic stem cells can be differentiated into multiple cell types, including cells in the cardiovascular systems. The appropriate cell types with the associated networks of vascular cells can be used to treat heart, lung, and blood diseases. This project is aiming at developing a novel systematic approach to understand, define, and ultimately control the process of stem cell differentiation, with the ultimate goal of developing tools of regenerative medicine to treat cardiovascular diseases.

描述（申请人提供）：多能人胚胎干细胞（hESC）可以在体外分化成多种细胞类型，包括心血管细胞（心肌细胞、血管内皮细胞和平滑肌细胞）。体外控制干细胞生长和分化的能力对于分化细胞成功应用于细胞疗法至关重要。适合所需谱系的细胞类型与心血管系统的相关元件一起可用于治疗心血管疾病。 所提出的确定控制 hESC 命运的最佳条件的研究使用了由综合化学物理微环境组成的新型中高通量微阵列平台。这些包括（1）固定化生长因子（GF）和细胞外基质蛋白（ECMP），（2）ECM的机械特性，以及（3）作用于细胞的外部机械力。目前的知识表明，这些参数（即 GF、ECMP、基质刚性以及机械负荷）中的每一个都在调节干细胞命运中发挥作用，但单独作用时效率较低。由于干细胞在发育阶段会受到多种微环境因素的影响，因此有必要研究当前研究中提出的生态位多因素复杂性的组合效应。在拟议的研究中，我们将研究 ECM 蛋白 (ECMP) 和 GF 对 hESC（联邦批准的 WA01 和 WA09 细胞系）分化的组合效应。我们为干细胞培养中的 ECMP 阵列设计了一种水凝胶系统来控制基质的刚性，涵盖不同组织中遇到的一系列刚性。我们还将把我们实验室开发的流动和拉伸装置整合到微阵列系统中，以评估外部机械力在 ECMP/GF/Rigidity 平台上调节 hESC 细胞命运的作用。这种新颖的组合微阵列系统可以对化学物理微环境进行全面测试，以选择 hESC 生长和分化的最佳条件。使用这种经过优化选择的hESCs转化为心血管组织工程将为显着促进人造血管、血管生成补片以及心力衰竭细胞替代的发展提供机会，从而改善心血管疾病患者的医疗保健以及我们公民的福祉。 公共健康相关性：多能人类胚胎干细胞可以分化成多种细胞类型，包括心血管系统细胞。具有相关血管细胞网络的适当细胞类型可用于治疗心脏、肺和血液疾病。该项目旨在开发一种新颖的系统方法来理解、定义和最终控制干细胞分化过程，最终目标是开发治疗心血管疾病的再生医学工具。