Cell Culture Core

核心B是中央细胞培养设施，将为四个项目中的每一个项目提供支持。该核心设施将提供以下服务：1)从小鼠血液和骨髓中分离中性粒细胞(PMN)；2)分离、鉴定和培养小鼠肺内皮细胞；以及3)培养人肺微血管和肺动脉内皮细胞，用于下文核心描述中概述的项目所涉及的实验。小鼠肺内皮细胞的分离和培养是完成项目1、2、3和4中提出的研究的关键。核心B还将从小鼠血液和骨髓中分离PMN，用于项目1、2和3。该核心的主要任务将是提供来自不同基因敲除小鼠的肺内皮细胞和PMN；例如，项目1的TRPM2基因敲除、NADPH氧化酶亚单位gp91(Phox)基因敲除和PKC-α基因敲除小鼠；项目2的MKP5基因敲除、P47(Phx)基因敲除、MK2基因敲除和MKP5/MK2-双基因敲除小鼠；TRPC4基因敲除；TRPM2基因敲除和Dream基因敲除小鼠；FoxMI转基因和iCKO小鼠用于项目4。因此，该核心的辐射能力对于项目研究的成功至关重要。

Core B is the central cell culture facility, which will provide support for each of four projects. This core facility will provide services for the following: 1) isolation of neutrophils (PMNs) from mouse blood and bone marrow; 2) isolation, characterization, and culture of mouse lung endothelial cells, and 3) culture of human lung microvascular and pulmonary artery endothelial cells for the experiments involved in the Projects as outlined in the Core description below. Mouse lung endothelial cell isolation and culture is pivotal for the accomplishment of studies proposed in Projects 1, 2, 3, and 4. In addition. Core B will also isolate PMNs from mouse blood and bone marrow for Projeds 1, 2, and 3. A major task of this core will be to provide lung endothelial cells and PMNs from different knock-out mice; e.g., TRPM2 knock-out, NADPH oxidase subunit gp91(phox) knock-out, and PKC-alpha knock-out mice for Project 1; MKP5 knock-out, p47(phox) knock-out, MK2 knock-out, and MKP5/MK2-double knock-out mice for Project 2; TRPC4 knock-out; TRPM2 knock-out, and Dream knock-out mice for Project 3; FoxMI transgenic and iCKO mice for Project 4. Thus the adivity of this Core is essential for the success of the Program Projed studies.