Core B is the Cell Culture and Small Animal Core. The function of the Cell Culture component is to provide Program Investigators with complete cell culture and cell isolation services that facilitate completion of their specific aims. In this capacity, the Core will: a) achieve economy of scale and quality assurance through centralized cell and tissue processing;b) isolate cells from the transgenic mice, including tau -/- (Project 1), PDE4B -/-, and PDE4D -/- (Project 2), and alG -/-, and eNOS -/- mice (Project 3);c) provide technical expertise, consultation, and training in cell isolation and culture;and d) serve as a centralized repository and source of endothelial cells and information generated from them. Considering that endothelial cells lining the pulmonary arteries, alveolar capillaries, and pulmonary veins exhibit remarkable heterogeneity both in structure and in function, the isolation and culture of endothelial cells from these different vascular segments is essential to the success of this application. Our Core routinely isolates pulmonary artery (PAEC), microvascular (PMVEC), and vein (PVEC) endothelial cells from transgenic and non-transgenic rats and mice. These cells are made available to investigators as needed. Specifically, we provide a seeding service, so that cell requests are filled twice per week, at the requested seeding density and in the requested tissue culture format. The Core currently maintains multiple cell lines for each aforementioned phenotype, seeds """"""""95 million cells per week, and provides cells to more than 50 laboratories nationally. The small animal component is a new constituent of the program. The function of the Small Animal component is to provide Program Investigators with a clinically relevant animal model of lung injury that facilitates completion of their Specific Aims. In this capacity, the core will: a) expand and maintain wild type and genetically modified strains of Pseudomonas aeruginosa;b) inject rats and mice with P. aeruginosa to elicit lung injury (a clinically relevant etiology of acute lung injury);c) monitor injected animals with P. aeruginosa until execution of terminal experiments;d) provide technical expertise, consultation and training in bacterial expansion and the animal model, and e) serve as a centralized repository of lung samples from infected and non-infected animals.
Isolation of endothelial cells Is fundamental to address the existing heterogeneity in signaling events proposed in the current application. The Cell Culture component of the Core centralizes isolation of the required endothelial cells, and provides the reagents necessary for completion of the proposed experiments in all projects. P. aeruginosa is a main clinical etiology of acute lung injury. The Small Animal component of the Core centralizes the development of a lung Injury model and provides training for the assessment of outcomes measures necessary for completion of the proposed experiments in all projects.
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