Core A: Cell Model and Evaluation Core

核心A：细胞模型和评估核心

• 批准号: 10246450
• 负责人: Bradford Alan Woodworth
• 金额: $ 24.17万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246450
• 关键词: Adopted; Anatomy; Animal Model; Area; Award; Biological Assay; Biology; Bronchi; Cell Culture System; Cell Culture Techniques; Cell model; Cells; Cellular Assay; Clinical Trials; Complement; Complex; Cost Savings; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Development; Disease; Dyes; Epithelial; Epithelial Cells; Equipment; Evaluation; Evolution; Fostering; Frequencies; Functional disorder; Funding; Gold; Health Insurance Portability and Accountability Act; Human; Human Resources; Hydration status; Image; In Situ; In Vitro; Individual; Institutional Review Boards; International; Intervention; Ion Transport; Ions; Laboratories; Laboratory Research; Lung Transplantation; Measurement; Measures; Messenger RNA; Methodology; Methods; Modeling; Morbidity - disease rate; Mucociliary Clearance; Mucous body substance; Nose; Operative Surgical Procedures; Optical Coherence Tomography; Pathogenesis; Pathway interactions; Permeability; Phenotype; Process; Property; Pulmonary Cystic Fibrosis; Reagent; Reproducibility; Research; Research Design; Research Personnel; Research Support; Resolution; Resources; Rights; Safety; Sampling; Science; Scientist; Source; Standardization; Structure of respiratory epithelium; Surface; System; Techniques; Technology; Testing; Thick; Tissues; Trachea; Training; Translating; Translations; United States National Institutes of Health; Validation; Viscosity; Western Blotting; Work; airway epithelium; airway inflammation; airway surface liquid; anatomic imaging; cystic fibrosis patients; digital; drug discovery; experience; human subject; improved; in vivo; innovation; insight; material transfer agreement; mortality; novel; novel therapeutics; protein expression; quality assurance; ranpirnase; real-time images; repository; response; sample fixation; standard measure

PROJECT SUMMARY / ABSTRACT: P30 CORE A Well-differentiated primary human airway epithelial cells and the assays that can be used with them are an instrumental model for understanding epithelial biology and are highly predictive of in vivo results in clinical trials. Primary cells can be used with methodologies that translate readily to assays of airway function in vivo, including measures of CFTR activity or other ion transporters, airway surface liquid depth and mucus hydration, and mucus viscosity and transport. Primary airway cells further provide an excellent model for examining the biology of airway epithelial inflammation, which is key to the pathogenesis of cystic fibrosis (CF) lung disease. The purpose of Core A is to support the research of numerous P30 investigators that involves cell culture systems and to assist with established and innovative assays available to characterize cellular responses. Core A carries out three main functions as outlined in the Specific Aims. First, Core A procures, grows, and distributes well-differentiated primary human airway epithelial cells from CF and non-CF donors. This includes samples of cells from lung transplants, surgically excised nasal tissue, and nasal brushings obtained at UAB and from a large array of collaborating centers. Examination of novel expansion and differentiation techniques for primary cells is also a key function of the Core. Second, Core A conducts functional anatomic imaging of airway epithelia by 1-micron resolution Optical Coherence Tomography (μOCT) in vitro (primary cells of human or non-human origin) and ex vivo (intact full-thickness trachea or mainstem bronchi of human origin or comparable tissues from CF animal models, thus interfacing with Core B). μOCT allows for investigators to explore mucus flow and mucociliary interactions and is designated as a National Core due to its unique and important capabilities. Third, Core A performs and assists with measures of CFTR activity and expression. In addition to traditional assays of CFTR function (e.g, Ussing chambers), the Core supports innovative conductance assays and advanced PCR technology for investigating CFTR and other protein expression. Core A facilitates interdisciplinary collaborative research, provides resources that are beyond the expertise of individual research laboratories, fosters the sharing of ideas and experimental strategies, assists with technical troubleshooting, and maintains essential equipment that is and will be heavily utilized by P30 personnel and beyond. Cost savings are achieved by minimizing duplicate efforts of individual CF investigators, by the centralized purchase and usage of equipment, reagents, and supplies, as well as by maintaining a central repository for human epithelial tissue. On the whole, Core A provides significant expertise and resources to aid P30 investigators, fostering advancement of epithelial cell culture and innovative assays to understand CFTR pathogenesis and support rational drug discovery.

项目摘要/摘要：P30核心A 分化良好的原代人呼吸道上皮细胞及其可用于检测的方法是 用于理解上皮生物学的仪器模型，并可高度预测临床中的活体结果 审判。原代细胞可以与易于转化为体内呼吸道功能分析的方法一起使用， 包括CFTR活性或其他离子转运体、呼吸道表面液体深度和粘液的测量 水合作用，粘液粘度和运输。原代呼吸道细胞进一步提供了一个极好的模型 检查呼吸道上皮炎症的生物学，这是囊性纤维化(CF)发病机制的关键 肺部疾病。核心A的目的是支持众多P30调查人员的研究，这些研究涉及 细胞培养系统，并协助已建立的和创新的可用于表征细胞的分析方法 回应。 核心A履行具体目标中概述的三项主要职能。首先，核心A获得、发展和 分化良好的原代人呼吸道上皮细胞来自CF型和非CF型供体。这包括 来自肺移植的细胞样本，手术切除的鼻组织，以及在UAB获得的鼻刷 以及来自一大批协作中心。新的扩展和分化技术的检验 对于原代细胞也是核心的一项关键功能。第二，核心A进行功能解剖成像 1微米分辨率光学相干断层扫描(μ-OCT)检测体外培养的人呼吸道上皮细胞 或非人类起源)和体外(完整的人类起源的全层气管或主干支气管或 来自CF动物模型的可比组织，因此与Core B接口)。μOCT允许调查人员 探索粘液流动和粘液纤毛相互作用，并被指定为国家核心，因为其独特的和 重要的能力。第三，核心A执行并协助测量CFTR活性和表达。在……里面 除了传统的CFTR功能分析(例如，Ussing小室)外，该核心还支持创新 电导分析和先进的聚合酶链式反应技术用于研究cftr和其他蛋白质的表达。 核心A促进跨学科协作研究，提供超越专业知识的资源 单独的研究实验室，促进思想和实验策略的分享，协助技术 故障排除，并维护P30人员和将大量使用的基本设备 更远一点。成本节约是通过最大限度地减少个别CF调查员的重复工作来实现的，由 设备、试剂和用品的集中采购和使用，以及通过维护中央 人类上皮组织的储存库。总体而言，核心A提供了重要的专业知识和资源 帮助P30研究人员，促进上皮细胞培养和创新分析的进步，以了解 Cftr的发病机制，支持合理的药物发现。