In Vitro and in Vivo Lung Studies

该细胞培养和动物核心旨在支持本项目中4个项目中每个项目的特定目标。 修改项目申请表。有两个具体目标。第一个目的是提供肺泡上皮细胞 来自人肺、大鼠肺和小鼠肺的II型细胞，以支持项目1的特定目标（Mostov）， 项目2（恩格尔）和项目3（罗森）。我们的研究小组在隔离， 培养和研究人和啮齿动物肺的肺泡上皮II型细胞， 正常和病理状态。这项工作在研究生物多样性的机制方面特别富有成效。 铜绿假单胞菌ExoT和ExoS对肺泡上皮细胞的损伤，以及确定ADP核糖基 铜绿假单胞菌ExoT的转移酶结构域有助于假单胞菌的生物活性 生物体（项目2，恩格尔）。此外，我们最近的工作已经确定，人类肺泡上皮细胞， II型细胞可以在基质胶中培养并诱导形成类似于真实的三维结构， 肺泡样结构。这一成就为肺的新研究提供了一个重要的机会 正常和病理条件下的上皮功能（项目1，Mostov）。这个核心的第二个目标， 代表了一个扩展的方向，是使用病理模型进行体内小鼠研究， 肺损伤，以检验项目3（罗森）和项目4（Werb）中的特定假设。这些体内小鼠 实验将使用感染性和非感染性刺激来测试对肺功能和整体的影响。 生存我们的研究小组在体内研究方面拥有丰富的经验， 在测试特定假设时产生的初步数据，项目3（罗森）的一部分， 项目4（Werb）。

This Cell Culture and Animal Core is designed to support specific aims in each of the 4 projects in this revised Program Project Application. There are two specific aims. The first aim is to supply alveolar epithelial type II cells from human lungs, rat lungs, and mouse lungs to support the specific aims of Project 1 (Mostov), Project 2 (Engel), and Project 3 (Rosen). Our research group has had extensive experience in isolating, culturing, and studying pulmonary alveolar epithelial type II cells from human and rodent lungs under both normal and pathological conditions. This work has been especially productive in studying mechanisms of alveolar epithelial injury from P.aeruginosa ExoT and ExoS as well as establishing how the ADP ribosyl transferase domain of P.aeruginosa ExoT contributes to the biologic activities of the Pseudomonas organisms (Project 2, Engel). In addition, our recent work has established that human alveolar epithelial type II cells can be cultured in matrigel and induced to form 3-dimentional structures that resemble authentic alveolar-like structures. This accomplishment provides a major opportunity for novel studies of lung epithelial function under normal and pathologic conditions (Project 1, Mostov). The second aim of this core, which represents an expanded direction, is to carry out in vivo mouse studies using pathologic models of lung injury to test specific hypotheses in Project 3 (Rosen) and Project 4 (Werb). These in vivo mouse experiments will use both infectious and non-infectious stimuli to test the effects on lung function and overall survival. Our research group has extensive experience with in vivo studies and there is considerable preliminary data that has been generated in testing specific hypotheses that a part of Project 3 (Rosen) and Project 4 (Werb).