Geometric Cues in the Stem Cell Microenvironment

干细胞微环境中的几何线索

• 批准号: 7891210
• 负责人: Kristopher Alan Kilian
• 金额: $ 5.05万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7891210
• 关键词: Adhesions; Adhesives; Adipocytes; Adult; Aspirate substance; Atherosclerosis; Biochemical; Biochemical Pathway; Biochemistry; Biological; Biological Assay; Bone Marrow; Cell Adhesion; Cell Fate Control; Cell Lineage; Cell Shape; Cell Therapy; Cells; Chemicals; Chemistry; Chondrocytes; Collection; Cues; Cytology; Cytoskeletal Modeling; DNA Microarray Chip; Differentiation and Growth; Disease; Electrophoresis; Elements; Event; Fatty acid glycerol esters; Focal Adhesions; Global Change; Histocytochemistry; Human; Immunofluorescence Immunologic; Lead; Masks; Mechanics; Mesenchymal Stem Cells; Microarray Analysis; Modeling; Monitor; Morphogenesis; Multipotent Stem Cells; Muscle; Muscle Cells; Neoplasm Metastasis; Neurons; Osteoblasts; Osteoporosis; Pathologic Processes; Pathology; Pathway interactions; Pattern; Physiological; Population; Printing; Protein Analysis; Proteins; RNA; RNA analysis; Regenerative Medicine; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Shapes; Signal Transduction; Source; Spatial Distribution; Staining method; Stains; Stem cells; Surface; System; Techniques; Tendon structure; Time; Tissue Engineering; Tissues; Variant; Western Blotting; Work; bone; cancer cell; cell type; density; design; enzyme activity; extracellular; improved; inhibitor/antagonist; insight; lipid biosynthesis; monolayer; multipotent cell; prevent; regenerative; regenerative therapy; self-renewal; small molecule; stem cell differentiation; stem cell fate; stem cell niche

DESCRIPTION (provided by applicant): The objective of this project is to explore how geometric cues in the niche guide mesenchymal stem cells (MSCs) towards specific fates. Human MSCs are multipotent stem cells that differentiate into fat, bone, tendon, muscle and neuron-like cells. The multipotency and ease of collection (i.e. fat aspirates, bone marrow) makes MSCs one of the more widely studied stem cell types for regenerative therapy and tissue engineering. Significant strides have been made in determining the chemical and biological signals that direct MSC fate decisions. However, there is currently very little understanding of the role of the niche geometry in directing MSC self-renewal and lineage commitment. The majority of systems that attempt to model the microenvironment of stem cells in their niche do not adequately control for variations in cell shape, spatial distribution and proliferation. The proposed research aims to use model surfaces to study the differentiation of an array of single MSCs confined to precise shapes. Cell adhesion and cytoskeletal features are important in many aspects of tissue morphogenesis. Thus, the choice of shapes for this work will be influenced by specific geometric cues previously shown to direct adhesive, contractile and protrusive elements. As the first specific aim, MSCs will be cultured on mixed patterns of shapes under growth and differentiation conditions to evaluate the effect of geometric cues by themselves and in combination with lineage promoting soluble factors. Histochemistry, immunofluorescence and RT-PCR will be used to compare cells across conditions and shapes. The second specific aim will explore inhibition of cytoskeletal elements and lineage regulators to isolate important biochemical pathways in shape-promoted fate decisions. Furthermore, the role of focal adhesion density within the niche will be explored in parallel by varying the surface chemistry across shapes. Along with techniques developed in aim 1, DNA microarray analysis, protein electrophoresis, western blotting and enzyme activity assays will be used to isolate important networks towards a thorough understanding of signaling events within the MSC niche. Mesenchymal stem cells (MSCs) from adult bone marrow hold promise as an abundant source of multipotent cells for regenerative medicine. Improper MSC fate regulation has been implicated in numerous pathologies including: osteoporosis, atherosclerosis and cancer cell metastasis. Therefore, deciphering how an adherent stem cells shape influences its fate is of vast importance for both understanding regenerative and pathological processes as well as for designing materials for stem cell based therapies.

描述（由申请人提供）：本项目的目的是探索生态位中的几何线索如何引导间充质干细胞（MSC）走向特定的命运。人骨髓间充质干细胞是多能干细胞，可分化为脂肪、骨、肌腱、肌肉和神经元样细胞。多能性和易于收集（即脂肪抽吸物，骨髓）使MSC成为再生治疗和组织工程中研究最广泛的干细胞类型之一。在确定指导MSC命运决定的化学和生物信号方面已经取得了重大进展。然而，目前很少有理解的作用，生态位的几何形状在指导MSC自我更新和谱系承诺。大多数试图模拟干细胞微环境的系统不能充分控制细胞形状、空间分布和增殖的变化。拟议的研究旨在使用模型表面来研究限于精确形状的单个MSC阵列的分化。细胞粘附和细胞骨架特征在组织形态发生的许多方面都很重要。因此，这项工作的形状的选择将受到特定的几何线索，以前显示直接粘合，收缩和膨胀元素的影响。作为第一个具体目标，MSC将在生长和分化条件下在混合形状模式上培养，以评估几何线索本身以及与谱系促进可溶性因子组合的效果。组织化学、免疫荧光和RT-PCR将用于比较不同条件和形状的细胞。第二个具体的目标将探讨抑制细胞骨架元素和谱系调节隔离重要的生化途径的形状促进命运的决定。此外，通过改变形状之间的表面化学性质，将平行探索壁龛内的粘着斑密度的作用。沿着在目标1中开发的技术，DNA微阵列分析、蛋白质电泳、蛋白质印迹和酶活性测定将用于分离重要网络，以彻底理解MSC生态位内的信号事件。来自成人骨髓的间充质干细胞（MSC）有望成为再生医学多能细胞的丰富来源。不适当的MSC命运调控已经涉及许多病理学，包括：骨质疏松症，动脉粥样硬化和癌细胞转移。因此，破译粘附的干细胞形状如何影响其命运对于理解再生和病理过程以及设计用于基于干细胞的疗法的材料都是非常重要的。