Core B is the Cell Culture and Small Animal Scientific Core whose goals are to provide Program Investigators with complete cell culture and cell isolation services (Cell Culture component) and a variety of small animal and microbiological services (Small Animal component) that facilitate the completion of their Specific Aims. To accomplish these goals, Core B will be centering its activities into three principal areas: Service component: Core B will facilitate the performance of each project by centralizing cell processing including isolation, characterization, and distribution of pulmonary endothelial cells (PAECs), microvascular endothelial cells (PMVECs), and pulmonary vein endothelial cells (PVECs) from wild type and genetically modified rats and mice, and by providing consultation and training in cell isolation and culture. Core B will facilitate the performance of each project by centralizing animal care including maintenance of mouse and rat colonies (breeding and colony expansion) of animals harboring genetic mutations, by maintaining bacterial stocks of Pseudomonas aeruginosa, and by generating P. aeruginosa-induced lung injury in rats and mice. Academic component: Throughout previous funding cycles, Core B has adapted to the ever-changing needs of the Projects by generating new reagents (e.g. providing pulmonary endothelial cells from genetically modified rats and mice) or platforms (e.g. providing an experimental rodent model of Pseudomonas aeruginosa-induced pneumonia and sepsis). During the current cycle, the Core has developed a novel experimental platform -Monolayer Stress Microscopy- to assess endothelial cell mechanics via visualization of forces occurring among cells and their subjacent substrate. In addition, the Core has generated mouse and rat decellularized lung bio-scaffolds that will serve as a platform for examining endothelial cell heterogeneity. Synergy with Projects and Scientific Cores: In collaboration with Core D (BioImaging and BioTechnology Implementation Core), pulmonary endothelial cells and isolated lung tissues will be provided for 3D, 4D, and 5D imaging analyses. Scientifically, the analysis of monolayer mechanical forces and the generation of lung scaffolds will become fundamental for data- and hypothesis- generation by providing innovative platforms to all Projects and Cores. In this capacity, scaffolds repopulated with genetically modified cells (provided from Core C) will be imaged using the novel approaches developed by Core D and the newly-acquired information will be disseminated in coordination with Core A (administrative Core).
Understanding the existing heterogeneity in the pulmonary circulation allowed the identification of disease mechanisms and the development of new therapeutic approaches to combat those illnesses. 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. 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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