Cell Culture Core

细胞培养核心

• 批准号: 7215381
• 负责人: Scott H Randell
• 金额: $ 12.35万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7215381
• 关键词: Acute; Air; Biochemical; Cell Separation; Cells; Clinical; Cultured Cells; Cystic Fibrosis Transmembrane Conductance Regulator; Diagnosis; Disease; Epithelial; Epithelial Cells; Foundations; Freezing; Genetic; Goals; Health; Human; Immunohistochemistry; In Situ Hybridization; Ion Transport; Liquid substance; Lung; Modeling; Mucins; Mucociliary Clearance; Mucous Membrane; Normal tissue morphology; North Carolina; Nose; Nucleosides; Nucleotides; Numbers; Output; Preparation; Purpose; Research; Research Personnel; Role; Source; Specimen; Support of Research; System; Tissue Procurements; Universities; airway surface liquid; demographics; disorder control; epithelial Na+ channel; in vivo; research study

Human airway epithelial cell cultures maintained at an air-liquid interface (ALI) recapitulate mucociliary differentiation found in vivo, regulating ion transport, mucin secetion and ciliated cell beating to reproduce mucociliary clearance mechanisms characterisitic of the native human mucosa. These cultures are an excellent model to study the role of CFTR, ENaC, nucleotide/nucleoside regulatory systems and their integrated function in health and disease as proposed in projects 1-4 of the current PPG, respectively. A Tissue Procurement and Cell Culture Core was established at the University of North Carolina in 1984, under the auspices of the CF Foundation, to provide standardized cell cultures to CF researchers. The Core has supported PPG projects since 2000 and has increased its output and capabilities to meet growing research demands. The Core routinely makes available cells and media that are unavailable and/or prohibitively expensive if purchased from commercial suppliers. The purpose of the present application is to provide continuity in the supply of airway specimens and large numbers of human airway epithelial cell cultures to PPG investigators, and to support the use of cultures in mechanistic studies. To accomplish these goals, we propose the following specific functions. 1) Tissue Procurement- Obtain nasal and bronchial tissues from normal, disease control, and CF humans as sources of airway epithelial cells and for acute ex vivo experiments, biochemical studies, in situ hybridization and immunohistochemistry. Characterize donors with respect to diagnosis, demographics, and clinical features relevant to research uses. 2) Cell Isolation and Culture- Isolate primary and passaged primary epithelial cells for distribution to project investigators. Maintain stocks of frozen cells and prepare media and substrates to support preparation of well-differentiated airway epithelial cultures as dictated by investigator needs. 3) Genetic Mainipulation of Cell Cultures- Provide support for adenoviral and retroviral expression of molecules under study by project investigators. Through these functions, and in conjunction with other Cores in this application, the Cell Culture Core will support research to better understand mechanisms underlying normal airway surface liquid homeostatsis, which is essential for healthy lungs.

人类气道上皮细胞培养物保持在空气界面（ALI）概括性粘膜纤毛 在体内发现的分化，调节离子传输，粘蛋白分解和纤毛细胞跳动以繁殖 人类粘膜的特征性粘膜钙清除机制。这些文化是 研究CFTR，ENAC，核苷酸/核苷调节系统及其其作用的出色模型 如当前PPG的1-4项目中提出的健康和疾病的综合功能。一个 组织采购和细胞培养核心于1984年在北卡罗来纳大学建立 在CF基金会的主持下，为CF研究人员提供标准化的细胞培养物。核心 自2000年以来一直支持PPG项目，并增加了其产出和能力来满足增长 研究要求。核心通常使不可用的细胞和媒体可用和/或 如果从商业供应商那里购买，则非常昂贵。本申请的目的是 提供气道标本和大量人类气道上皮细胞的连续性 向PPG研究者进行培养，并支持在机械研究中使用培养物。完成 这些目标，我们提出以下特定功能。 1）组织采购 - 获得鼻和支气管 正常，疾病控制和CF人类的组织作为气道上皮细胞的来源，急性EX 体内实验，生化研究，原位杂交和免疫组织化学。捐助者的特征 关于诊断，人口统计和与研究用途相关的临床特征。 2）细胞分离 和培养物分离的原发性和传递的原发性上皮细胞，以分布到项目研究者。 维护冷冻细胞的库存，并准备培养基和底物以支持良好差异的准备 气道上皮文化是由研究者需求决定的。 3）细胞培养物的遗传主要注入 - 为项目研究者研究的分子的腺病毒和逆转录病毒表达提供支持。 通过这些功能，并在此应用中与其他核心结合，细胞培养核心将 支持研究以更好地理解正常气道表面液体体内平衡的机制， 这对于健康的肺部至关重要。