UAB CFRC Core A: Cell Model and Evaluation Core

项目总结/摘要：P30核心A 分化良好的人气道上皮细胞、模型系统和可用于它们的测定 是理解上皮生物学的工具模型，并且高度预测体内结果， 临床试验原代细胞可以与容易转化为气道和呼吸道的测定的方法一起使用。 体内上皮功能，包括CFTR活性或其他离子转运蛋白的测量，气道表面液体 深度和粘液水合作用，粘液粘度和运输。上皮细胞模型进一步提供了一种极好的 用于检查气道上皮炎症的生物学的模型，这是囊性变发病机制的关键。 纤维化（CF）肺疾病。核心A的目的是支持众多P30研究者的研究 这涉及细胞培养系统，并协助建立和创新的分析，可用于表征 细胞反应。 核心A执行具体目标中概述的三个主要功能。首先，核心A（通过交互）获取 用于通过核心C）临床采集样品，生长和分布分化良好的原代人 来自各种组织来源（肺、窦、直肠等）的上皮细胞来自CF和非CF捐助者。这 包括来自肺移植的细胞样本，手术切除的鼻组织，直肠活检， 在UAB和从大量的合作中心获得的刷。审查新的扩展 原代细胞的分化技术也是核心的关键功能。第二，核心A进行 通过1微米分辨率光学相干断层扫描（μOCT）进行气道上皮功能解剖成像 体外（人或非人来源的原代细胞）和离体（完整的全层气管，主干 支气管和其他人源组织或来自CF动物模型的相当组织，从而与 核心B）。μOCT允许研究者探索粘液流动和粘膜纤毛相互作用， 作为一个国家的核心，由于其独特的和重要的能力。第三，核心A执行并协助 CFTR活性和表达的测量。除了CFTR功能的传统测定（例如，Ussing Core支持创新的电导测定、类器官肿胀测定和先进的PCR 研究CFTR和其他蛋白表达的技术。 核心A促进跨学科的合作研究，提供超出专业知识的资源， 个人研究实验室，促进思想和实验策略的共享，协助技术 排除故障，维护P30人员大量使用的必要设备， 超越。通过最大限度地减少个别CF调查员的重复工作， 集中采购和使用设备、试剂和供应品，以及维持一个 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检测的进步，以了解 CFTR发病机制和支持合理的药物发现。

PROJECT SUMMARY / ABSTRACT: P30 CORE A Well-differentiated human airway epithelial cells, model systems, 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 and epithelilal function in vivo, including measures of CFTR activity or other ion transporters, airway surface liquid depth and mucus hydration, mucus viscosity and transport. epithelial cell models 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 (via interactions for clinical acquisition of samples via Core C), grows, and distributes well-differentiated primary human epithelial cells from a variety of tissue sources (lung, sinus, rectal, etc.) from CF and non-CF donors. This includes samples of cells from lung transplants, surgically excised nasal tissue, rectal biopsies, 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, mainstem bronchi, and other tissues 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, organoid swelling 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.