Cell Culture and Vector Core

细胞培养和载体核心

• 批准号: 7457951
• 负责人: CHINNASWAMY TIRUPPATHI
• 金额: $ 24.23万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7457951
• 关键词: Adenoviruses; Blood; Blood Vessels; Cell Separation; Core Facility; Cultured Cells; Endothelial Cells; Human; Infection; Knock-out; Knockout Mice; Lung; Mus; NADPH Oxidase; Preparation; Recombinants; Services; TLR4 gene; Viral; human AKAP13 protein; neutrophil; programs; protein kinase C zeta; pulmonary artery endothelial cell; research study; vector

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.

核心B是中央细胞培养和载体设施，将为四个项目提供支持。该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构建体; 5）扩增重组腺病毒。 腺病毒构建体，和6）用重组腺病毒感染原代内皮细胞，用于下面核心描述中概述的项目中涉及的实验。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类和小鼠血液中分离中性粒细胞。该核心的主要任务是为项目1、3和4提供病毒表达构建体。该核心还将为项目2中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敲除小鼠的内皮细胞是关键的;例如，NADPH氧化酶（gp 91和p47敲除），项目1和4的PKC-zeta敲除小鼠，项目3的TRPC 4敲除小鼠，以及TLR 4 4号项目的基因敲除小鼠该核心的活动对于计划项目的成功至关重要。