CORE--CELL CULTURE CORE

核心--细胞培养核心

• 批准号: 6724366
• 负责人: Lou Ann S Brown
• 金额: $ 15.9万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6724366
• 关键词: adult respiratory distress syndrome; alcoholism /alcohol abuse; alveolar macrophages; biomedical facility; fibroblasts; glutathione; lung injury; respiratory epithelium; tissue /cell culture

DESCRIPTION (provided by applicant): The Acute Respiratory Distress Syndrome (ARDS) is a common form of acute lung injury with no effective therapy and mortality of 50%. Investigators in this Center application demonstrated that chronic alcohol abuse was the first co-morbid variable to be identified that significantly increases the incidence and severity of ARDS. In this Center application, the mechanisms by which chronic ethanol ingestion predisposes to sepsis-induced acute lung injury will be addressed in alveolar endothelial cells, alveolar epithelial cells, pulmonary fibroblasts, and alveolar macrophages isolated from a rat model of chronic ethanol ingestion. When the role of glutathione availability in ethanol-induced toxicity is the study question, the rats will be fed the glutathione precursors N-acetylcysteine or Procysteine (L-2-oxo-4-thiazolidine-carboxylic acid) either during chronic ethanol ingestion. The purpose of the cell culture core will be to isolate and culture these cells after the appropriate dietary regimen and then distribute the cells to the different projects. For Projects 1 and 2 and Pilot Project 2, the cell culture core will provide primary alveolar type II cells. For Project 1, alveolar macrophages will also be provided. For Project 3 and Pilot Project 2, the core will provide pulmonary fibroblasts and alveolar type II cells from control or ethanol-exposed rats. For Projects 4 and 5 and Pilot Project 2, the core will be responsible for the isolation and culture of pulmonary microvascular endothelial cells. Peripheral neutrophils will also be provided for Project 4. This core will provide cells with maximum reproducibility, efficiency and cost effectiveness. These primary cells will be isolated and cultured in strict endotoxin-free conditions by experienced cell culture technicians. The purity of the cells obtained will routinely be verified using immunohistochemical techniques and functional assays. Cultured cells will be distributed to all investigators according to their requests for studies outlined in their proposals.

描述（由申请人提供）：急性呼吸窘迫综合征（ARDS）是 急性肺损伤的常见形式，没有有效的治疗和50％的死亡率。该中心申请中的研究人员表明，慢性酒精滥用是第一个被确定可显着增加ARDS的发病率和严重性的合并变量。在此中心应用中，将在肺泡内皮细胞，肺泡上皮细胞，肺成纤维细胞以及从慢性乙醇摄入大鼠模型中分离出的肺泡内皮细胞，肺泡成纤维细胞和肺泡巨噬细胞的慢性乙醇诱发急性肺损伤的机制。当研究问题是谷胱甘肽在乙醇引起的毒性中的作用时，将在慢性乙醇摄入期间给大鼠喂食大鼠N-乙酰半胱氨酸或procySteine（L-2-oxo-4-噻唑烷二羧酸）。细胞培养核心的目的是在适当的饮食方案后分离和培养这些细胞，然后将细胞分配给不同的项目。对于项目1和2和试点项目2，细胞培养核心将提供原发性牙槽II型细胞。对于项目1，还将提供肺泡巨噬细胞。对于项目3和试点项目2，核心将通过对照或暴露于乙醇的大鼠提供肺成纤维细胞和肺泡II型细胞。对于项目4和5和试点项目2，核心将负责肺微血管内皮细胞的隔离和培养。项目4也将提供外围性嗜中性粒细胞。此核心将 为细胞提供最大的可重复性，效率和成本效益。这些原代细胞将通过经验丰富的细胞培养技术人员在严格的无内毒素条件下分离和培养。通过免疫组织化学技术和功能测定，将常规验证获得的细胞的纯度。培养的细胞将根据所有研究人员的提议中概述的研究要求分配给所有研究者。