Live Imaging of Intact Airway

完整气道的实时成像

• 批准号: 10240652
• 负责人: Charles P. Lin
• 金额: $ 55.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240652
• 关键词: 3-Dimensional; Ablation; Advanced Development; Affect; Airway Disease; Basal Cell; Behavior; Binding; Biology; Biomechanics; Biomedical Engineering; Blood Vessels; Cell Differentiation process; Cell Proliferation; Cell physiology; Cells; Cilia; Complex; Data; Development; Disease; Engraftment; Epithelial; Epithelial Cells; Equilibrium; Event; Extracellular Matrix; Genetic; Heterogeneity; Homeostasis; Human; Image; Imaging Techniques; Imaging technology; Individual; Laboratories; Laser Scanning Microscopy; Lasers; Learning; Lung diseases; Maintenance; Mechanics; Mediating; Mesenchymal; Mesenchyme; Migration Assay; Minor; Modeling; Modification; Motion; Mucociliary Clearance; Mus; Natural regeneration; Organ; Organ Culture Techniques; PDGFRB gene; Periodicity; Physiological; Play; Population; Process; Public Health; Reporter; Resolution; Respiratory System; Role; Signal Transduction; Specific qualifier value; Stretching; Stromal Cells; Supporting Cell; System; Testing; Time; Tissues; Toxin; Transplantation; Visualization; Work; airway epithelium; airway regeneration; behavior test; cell behavior; cell motility; cell type; design; epithelial stem cell; epithelium regeneration; experimental study; fluid flow; imaging modality; imaging platform; individual response; injured airway; intraepithelial; mechanical force; migration; multiphoton imaging; novel; optical imaging; prevent; response to injury; scaffold; shear stress; stem; stem cell biology; stem cell differentiation; stem cell function; stem cell niche; stem cells; tissue culture; two-photon

The integrity of the lining of the airways relies on multiple functions of its epithelial stem cells. We hypothesize that the airway stem cell behavior is regulated by the cells and the forces that surrounds them, including other epithelial cells, subepithelial mesenchymal cells, and external mechanical forces exerted on the epithelium. How these varied components regulate the behavior of airway progenitor cells is largely unknown. Furthermore, basal stem cell – niche interactions may explain the increasingly recognized heterogeneity within this progenitor cell population, including regional and tissue-level heterogeneity in basal stem cell behavior. Although progenitor cell – niche interactions have been described with live imaging in other systems, the constant motion and challenging access of the respiratory system has hampered the development of platforms for live imaging of the airways at high resolution. We combined emerging stem cell biology and live imaging technologies to develop a novel airway explant live imaging platform. We imaged the maintenance and regeneration of the airway epithelium in a mouse airway explant, surrounded by all of the major components of its 3D multicomponent microenvironment, including the ECM and the mesenchyme. We present preliminary data on tracking individual stem cells during regeneration with two photon laser scanning microscopy, including visualization of stem cell differentiation with fluorescent reporters and analysis of ciliated cell function with novel video-rate imaging modalities. To define the airway stem cell microenvironment and to test its role in progenitor cells function, we visualize and putative cellular niche components and test their role by laser and toxin-mediated ablation. We propose to use cellular engraftment followed by application of mechanical forces to check the role of the microenvironment in establishing planar polarity, and to study the relationship of proliferation, migration and differentiation in collective cell migration in the airway epithelium.

气道内壁的完整性依赖于其上皮干细胞的多个功能。 我们假设气道干细胞行为受细胞和力调节 包围它们，包括其他上皮细胞，地下间质细胞和外部 机械力施加在上皮上。这些不同的组件如何调节 气道祖细胞的行为在很大程度上未知。此外，基本干细胞 - 利基 相互作用可以解释该祖细胞中越来越公认的异质性 种群，包括基本干细胞行为中的区域和组织水平的异质性。 虽然祖细胞 - 在其他现场成像中已经描述了利基相互作用 系统，呼吸系统的持续运动和挑战访问受到阻碍 高分辨率以高分辨率的实时成像的平台开发。 我们结合了新兴的干细胞生物学和实时成像技术，以开发一种新颖的 Airway Explant Live Imaging平台。我们成像了维护和再生 鼠标气道外植体中的气道上皮，周围是其所有主要组成部分 3D多组分微环境，包括ECM和间质。我们在场 使用两个光子激光再生过程中跟踪单个干细胞的初步数据 扫描显微镜，包括可视化干细胞分化与荧光 记者和新型视频速率成像方式的纤毛细胞功能分析。到 定义气道干细胞微环境并测试其在祖细胞功能中的作用，我们 可视化和推定的细胞小众组件，并通过激光和毒素介导的角色测试其作用 消融。我们建议使用细胞植入，然后将机械力应用于 检查微环境在建立平面极性中的作用，并研究 气道集体细胞迁移中增殖，迁移和分化的关系 上皮。