Core B is the central cell culture facility, which provides support for each of the three projects. The cell culture Core will perform the following functions and services: 1) isolate and culture alveolar and bone marrow-derived macrophages from wild type and genetically modified mice; 2) isolate and culture macrophages from human bronchoalveolar fluid; 3) isolate monocytes from human pheripheral blood; 4) Maintenance and propagation of primary human lung microvessel endothelial cells; 5) isolate, culture, and characterize lung endothelial cells from wild type and genetically modified mice; 6) isolate, culture, and characterize lung epithelial (Type I and Type II) cells from wild type and genetically modified mice; and 7) provide technical support for gene transcription regulation studies. Core B will supply alveolar and bone marrow- derived macrophages from wild type and knockout (Trpm2?/?, Nox2?/?, Nox4?/?, Cd38?/?, Trpm2Mye?/?, Nlrp3?/?, and caspase1?/?) mice, and human alveolar macrophages for the accomplishment of studies proposed in Project 1. For Project 2 studies, Core B will provide mouse lung endothelial and Type II epithelial cells, and supply human epithelial cells. In addition, Core B will also provide for studies in Project 2 bone marrow-derived macrophages from WT, Sphk2?/?, Par?/?, and Creb1Mye?/? mice. For studies proposed in Project 3, Core B will provide human monocytes and lung endothelial cells, lung endothelial cells from WT and Jag1i?EC mice, and alveolar and bone marrow-derived macrophages from wild type and knockout (Jag1i?EC, Notch1?Mye, Notch2?Mye, and Notch1-Notch2?Mye) mice. Additionally, Core B will provide technical support for analysis of gene transcription studies proposed in Projects 2 and 3. This Core is essential for the success of the Program Project.
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