Engineering 3D in vitro niches to reveal fundamentals of cellular biomechanics

工程 3D 体外利基揭示细胞生物力学的基础知识

• 批准号: 7849439
• 负责人: Sarah C Heilshorn
• 金额: $ 240万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7849439
• 关键词: Address; Adult; Biochemical; Biocompatible Materials; Biological; Biological Models; Biomechanics; Biomedical Research; Brain; Cell Culture Techniques; Cells; Cellular biology; Chemotaxis; Cues; Development; Disease; Engineering; Future; In Vitro; Injury; Ligands; Mammalian Cell; Medicine; Methods; Microfluidics; Muscle Rigidity; Natural regeneration; Neuraxis; Pattern; Protein Engineering; Protocols documentation; Resolution; Scientist; Signal Transduction; Site; Solutions; Technology; Testing; Tissue Culture Techniques; abstracting; cell motility; cell type; in vivo; innovation; nerve stem cell; public health relevance

DESCRIPTION (Provided by the applicant) Abstract: The development of tissue culture techniques by Ross Granville Harrison in 1907 has been cited as one of the ten greatest discoveries in medicine and enabled monumental advances in biological understanding. Despite the enduring importance of in vitro culture in modern biomedicine, the technology of mammalian cell culture has remained largely unchanged since the 1940's: cells are cultured on hard, flat substrates and surrounded by homogeneous solutions of medium that do little to recreate the exquisite microenvironments found in vivo. Cells are well known to respond to multiple cues found within their in vivo niches, e.g., concentration gradients of soluble and tethered biochemicals, matrix rigidity, patterns of matrix ligands, and interactions with other cell types; however, few methods exist to recapitulate these cues in in vitro cell culture studies. To address these limitations, I propose creating versatile, three-dimensional in vitro niches with precise spatial and temporal resolution of cellular cues. These three-dimensional microenvironments will be fabricated using innovative and transdisciplinary approaches that combine advances in protein engineering, biomaterials, and microfluidics with traditional cell biology protocols. As a model system, these in vitro niches will be used to quantitatively study the cellular biomechanics and signaling mechanisms regulating neural progenitor cell (NPC) migration. NPC chemotaxis within gradients of soluble factors is hypothesized to be contextual and reliant on additional biomechanical cues from the 3D matrix. The presence of NPCs within specific niches of the brain opens up the tantalizing possibility that the adult central nervous system may be able to regenerate following injury or disease if NPCs were induced to migrate to sites of need. The development of quantitative, in vitro mimics of in vivo niches will have a profound impact on biomedical research by enabling scientists to test entirely new hypotheses about the interactions between different cells and their three-dimensional microenvironments. Public Health Relevance: Despite the enduring importance of tissue culture techniques in modern biomedicine, the technology of mammalian cell culture has remained largely unchanged since the 1940's: cells are cultured on hard, flat substrates and surrounded by solutions of medium that do little to recreate the exquisite microenvironments (called niches) found inside the body. To address these limitations, I propose creating versatile mimics of three-dimensional niches with precise spatial and temporal resolution of cellular cues. As a model system, these engineered niches will be used to quantitatively study the cellular biomechanics and signaling mechanisms regulating neural progenitor cell (NPC) migration, opening the door to future therapies for regeneration of the central nervous system.

描述（由申请人提供） 摘要：1907年罗斯·格兰维尔·哈里森（Ross Granville Harrison）开发的组织培养技术被列为医学十大发现之一，并使生物学理解取得了巨大进步。尽管体外培养在现代生物医学中具有持久的重要性，但自20世纪40年代以来，哺乳动物细胞培养技术基本上保持不变：细胞在坚硬平坦的基底上培养，并被均匀的培养基溶液包围，这些溶液几乎无法重现体内发现的精致微环境。众所周知，细胞对在其体内小生境中发现的多种线索作出反应，例如，浓度梯度的可溶性和拴系的生化物质，基质刚性，模式的基质配体，和与其他细胞类型的相互作用;然而，很少有方法存在，以概括这些线索在体外细胞培养研究。为了解决这些局限性，我建议创建通用的，三维的体外壁龛精确的空间和时间分辨率的细胞线索。这些三维微环境将使用创新和跨学科的方法制造，这些方法将联合收割机在蛋白质工程，生物材料和微流体方面的进展与传统的细胞生物学协议相结合。作为一个模型系统，这些在体外壁龛将被用来定量研究细胞的生物力学和信号机制调节神经祖细胞（NPC）的迁移。假设可溶性因子梯度内的NPC趋化性是上下文相关的，并且依赖于来自3D矩阵的额外生物力学线索。在大脑的特定壁龛中存在NPC开辟了一种诱人的可能性，即如果NPC被诱导迁移到需要的部位，成年中枢神经系统可能能够在损伤或疾病后再生。体内小生境的定量体外模拟物的开发将对生物医学研究产生深远的影响，使科学家能够测试关于不同细胞及其三维微环境之间相互作用的全新假设。 公共卫生相关性：尽管组织培养技术在现代生物医学中具有持久的重要性，但自20世纪40年代以来，哺乳动物细胞培养技术在很大程度上保持不变：细胞在坚硬平坦的基质上培养，并被培养基溶液包围，这些溶液几乎无法重建体内发现的精致微环境（称为壁龛）。为了解决这些局限性，我建议创建多功能的模仿三维壁龛精确的空间和时间分辨率的细胞线索。作为一个模型系统，这些工程小生境将用于定量研究调节神经祖细胞（NPC）迁移的细胞生物力学和信号机制，为未来的中枢神经系统再生疗法打开大门。