The Microenvironment Effects on the Osteogenesis of Human Embryonic Stem Cells

微环境对人胚胎干细胞成骨的影响

• 批准号: 7764657
• 负责人: Shelley Elaine Brown
• 金额: $ 3.7万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7764657
• 关键词: Adipocytes; Adult; Affect; Allografting; Architecture; Area; Biocompatible Materials; Biology; Bone Marrow; Bone Tissue; Calvaria; Cell Culture Techniques; Cell Differentiation process; Cells; Chondrocytes; Coculture Techniques; Collagen; Defect; Deposition; Development; Embryo; Encapsulated; Environment; Extracellular Matrix; Fluorescence-Activated Cell Sorting; Gel; Gene Expression; Goals; Growth Factor; Human; Implant; In Vitro; Intramuscular; Knowledge; Measures; Mesenchymal; Mesenchymal Stem Cells; Methods; Microarray Analysis; Modeling; Molecular Profiling; Organogenesis; Osteoblasts; Osteogenesis; Permeability; Phase; Population; Rattus; Regenerative Medicine; Replacement Therapy; Research; Scanning; Signal Transduction; Solid; Stem Cell Development; Stem cell transplant; Stromal Cells; Subfamily lentivirinae; System; Techniques; Time; Tissue Engineering; Tissue-Specific Gene Expression; Tissues; Transgenes; Transplantation; Work; Wound Healing; X-Ray Computed Tomography; base; bone; cell type; craniofacial repair; design; human embryonic stem cell; in vivo; interest; osteoblast differentiation; osteogenic; osteoprogenitor cell; public health relevance; repaired; repository; scaffold; skeletal; tissue regeneration

DESCRIPTION (provided by applicant): Human embryonic stem cells (hESCs) present a potentially unlimited supply of cells that may be directed to differentiate into all cell types within the body and used in regenerative medicine for tissue and cell replacement therapies. An area of particular interest is stem cell transplantation for bone tissue regeneration where hESCs may be used to repair skeletal defects. Current techniques used for bone tissue repair employ the use of auto- and allografting methods, however, these methods have inherent limitations that restrict their universal application. The limitations of these reparative strategies suggest that an alternative approach is required, and hESCs may provide a repository of cells for such an approach. One of the major gaps in the knowledge regarding hESCs is the lack of understanding of the growth factors and three-dimensional signals that control differentiation. Previous work has demonstrated that hESCs can be directed to osteoblasts, however, how to achieve directed differentiation still remains a pivotal question that is unanswered. Therefore, we hypothesize that controlling the local in vitro and in vivo microenvironments can control osteoblast differentiation of hESCs. Here, we investigate the importance of cell culture conditions in-the in vitro microenvironment, and the importance of scaffold design in the in vivo microenvironment. We will begin by developing two hESC co-culture systems to direct osteoblast differentiation via pro-osteogenic factors derived from both hBMSCs and mesenchymal cells derived from hESCs (ES-MSCs). Then, we will identify a homogenous population of osteoprogenitor hESCs using the lineage specific Col2.3-GFP transgene, fluorescence-activated cell sorting (FACS) and microarray analysis. Next, we will deliver the osteoprogenitor hESCs in vivo within scaffolds to investigate the effects pro-osteogenic factor incorporation and scaffold permeability have on cell differentiation and overall bone tissue formation via micro-CT. PUBLIC HEALTH RELEVANCE: The long-term goal of this research is to further understand the biology of human embryonic stem cell development and more specifically, provide information about the organogenesis of bone. Through this research, I hope to contribute to the biomedical field in an effort to aid in the development of new hESC-based tissue engineering and regenerative medicine strategies.

描述（由申请人提供）：人类胚胎干细胞（hESC）提供了潜在无限的细胞供应，可定向分化为体内所有细胞类型，并用于组织和细胞替代疗法的再生医学。一个特别令人感兴趣的领域是用于骨组织再生的干细胞移植，其中 hESC 可用于修复骨骼缺陷。目前用于骨组织修复的技术采用自体和同种异体移植方法，然而，这些方法具有限制其普遍应用的固有局限性。这些修复策略的局限性表明需要一种替代方法，而 hESC 可以为这种方法提供细胞库。关于 hESC 知识的主要差距之一是缺乏对控制分化的生长因子和三维信号的了解。先前的工作已经证明hESCs可以定向成骨细胞，然而，如何实现定向分化仍然是一个尚未解答的关键问题。因此，我们假设控制局部体外和体内微环境可以控制hESCs的成骨细胞分化。在这里，我们研究了体外微环境中细胞培养条件的重要性，以及体内微环境中支架设计的重要性。我们将首先开发两种 hESC 共培养系统，通过 hBMSC 和 hESC 间充质细胞 (ES-MSC) 衍生的促成骨因子指导成骨细胞分化。然后，我们将使用谱系特异性 Col2.3-GFP 转基因、荧光激活细胞分选 (FACS) 和微阵列分析来鉴定同质骨祖细胞 hESC 群体。接下来，我们将在支架内体内递送骨祖细胞 hESC，通过微 CT 研究促成骨因子掺入和支架通透性对细胞分化和整体骨组织形成的影响。公共健康相关性：这项研究的长期目标是进一步了解人类胚胎干细胞发育的生物学，更具体地说，提供有关骨骼器官发生的信息。通过这项研究，我希望为生物医学领域做出贡献，帮助开发基于 hESC 的新组织工程和再生医学策略。