Cell Culture and Vector Core

核心B是中央细胞培养和载体设施，将为四个项目中的每一个项目提供支持。这一核心设施将提供以下服务：1)从人和小鼠血液中分离中性粒细胞；2)分离、鉴定和培养小鼠肺内皮细胞；3)培养人肺微血管和肺动脉内皮细胞；4)制备重组腺病毒表达载体；5)扩增重组人肺内皮细胞。 腺病毒构建，以及6)用重组腺病毒感染原代内皮细胞，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、3和4中提出的研究的关键。此外，这三个项目将需要人肺微血管和肺动脉内皮细胞。该核心还将为所有项目从人和鼠的血液中分离中性粒细胞。这个核心的主要任务将是为项目1、3和4提供病毒表达结构。该核心还将为项目2的研究提供中性粒细胞和内皮细胞。此外，该核心对于提供来自不同基因敲除小鼠的内皮细胞至关重要；例如，NADPH氧化酶(gp91和p47基因敲除)、项目1和4的PKC-Zeta基因敲除小鼠、项目3的TRPC4基因敲除小鼠和TLR4 项目4的淘汰赛老鼠。这个核心的活动对项目的成功至关重要。

Core B is the central cell culture and vector facility, which will provide support for each of four projects. This core facility will provide services for the following: 1) isolation of neutrophils from human and mouse blood; 2) isolation , charcterization, and culture of mouse lung endothelial cells; 3) culture of human lung micro vascular and pulmonary artery endothelial cells; 4) preparation of recombinant adenovirus expression constructs; 5) amplifying recombinant adenovirus constructs, and 6) infection of primary endothelial cells with recombinant adenoviruses 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 1,3, and 4. In addition, these three projects will require human lung microvascular and pulmonary artery endothelial cells. The core will also isolate neutrophils from human and mouse blood for all projects. A major task of this core will be to provide viral expression constructs for the Projects 1,3, and 4. This core will also provide neutrophils and endothelial cells for the studies in Project 2. In addition, this Core is critical for providing endothelial cells from different knockout mice; e.g., NADPH oxidase (gp91 and p47 knockout), PKC-zeta knockout mice for Projects 1 and 4, TRPC4 knockout mice for Project 3, and TLR4 knockout mice for Project 4. The activity of this Core is essential for the successs of the Program Project.