The Cell Phenotyping and Mouse Core

细胞表型和小鼠核心

基本信息

批准号：
10696959
负责人：
Alexander Misharin
金额：
$ 36.65万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10696959
关键词：
Acceleration Alveolar Alveolar Macrophages Animal Experiments Animal Model Animals Attention B-Lymphocytes Blinded Blood Vessels Bone Marrow Breeding Cell Separation Cells Chest Chimera organism Collagen Cre driver DNA Dendritic Cells Deposition Endothelial Cells Enterobacteria phage P1 Cre recombinase Environment Epithelial Cells Epithelium Fibroblasts Flow Cytometry Funding Opportunities Generations Genes Genetically Engineered Mouse Genotype Histologic Human Resources Immune Individual Infection Inflammatory Influenza Influenza A Virus, H1N1 Subtype Influenza A virus Instruction Intervention Knock-out Knockout Mice Libraries Liquid substance Lung Lung Compliance Macrophage Measurement Measures Mechanics Modeling Molecular Morbidity - disease rate Mus Natural Killer Cells Outcome Measure Pathway interactions Phenotype Pneumonia Population Procedures Proteins Pulmonary Inflammation RNA Recovery Reporter Reproducibility Research Research Personnel Resistance Resources Sorting Standardization Strategic Planning Structure System T-Lymphocyte T-Lymphocyte Subsets Techniques Testing Tissues Titrations Transgenic Organisms Type II Epithelial Receptor Cell Viral Viral Pneumonia Virus aged alveolar epithelium cell type central database conditional knockout cost cytokine eosinophil epigenomics experience experimental study indexing inducible Cre influenza A pneumonia interest interstitial lung injury lung repair lymphoid organ mast cell metabolomics monocyte mortality mouse model neutrophil non-invasive imaging overexpression programs recruit repaired response second harmonic single-cell RNA sequencing synergism tool transcriptome sequencing transcriptomics

项目摘要

Project Summary The Cell Phenotyping and Mouse Core, designated as Core C, is a centralized facility that will provide investigators with a well-established and reproducible model of influenza A–induced pneumonia. Core C will generate stocks of influenza (A/WSN/33) virus, titrate and maintain viral stocks, and perform infection of the experimental animals as outlined in Projects 1, 2, 3 and 4. Quantitative tools such as Flexivent measurements of lung mechanics, quantitative histological assessment, flow cytometric phenotyping and sorting of the immune cellular populations, and multiplex measures of pro-inflammatory cytokines will be conducted by Core C according to the research plan outlined in Projects 1, 2, 3 and 4. Core C will use advanced flow cytometry capabilities to quantify, phenotype, and sort specific inflammatory and parenchymal cell populations (such as tissue-resident and recruited alveolar macrophages, perivascular and peribronchial interstitial macrophages, monocyte subsets, dendritic cell subsets, T, B and NK cell subsets, alveolar epithelial type II cells, fibroblast subsets, endothelial cells) from the murine lung and lymphoid organs. The sorting and isolation capabilities of the Core will allow cell-type-specific assessment of transcriptomic response via RNA-sequencing, including single-cell RNA-sequencing. To this matter, Core will provide automated DNA/RNA isolation from sorted cells, followed by quality assessment and RNA-sequencing library construction and sequencing. Core C will breed mice to generate the cell-type- or tissue-specific Cre recombinase lines to induce tissue-specific knockout mice. Core C will perform the genotyping of all the murine strains proposed by Project Leaders. Core C will maintain a uniform environment in which wild-type and genetically engineered mice will be maintained, subjected to lung injury and allowed to recover. Collectively, these tools provide a unique resource for Project Investigators, which would be difficult to reproduce without the support of this PPG.

项目概要细胞表型分析和小鼠核心，指定为核心 C，是一个集中设施，将提供拥有完善且可重复的甲型流感诱发的核心 C 肺炎模型的研究人员将会这样做。产生流感 (A/WSN/33) 病毒库存，滴定并维持病毒库存，并进行感染项目 1、2、3 和 4 中概述的实验动物。定量工具，例如 Flexivent 测量肺力学、定量组织学评估、流式细胞术表型分析和分类免疫细胞群和促炎细胞因子的多重测量将由 Core 进行 C 根据项目 1、2、3 和 4 中概述的研究计划。核心 C 将使用先进的流式细胞术量化、表型和分类特定炎症和实质细胞群的能力（例如组织驻留和招募的肺泡巨噬细胞、血管周围和支气管周围间质巨噬细胞，单核细胞亚群、树突状细胞亚群、T、B 和 NK 细胞亚群、肺泡上皮 II 型细胞、成纤维细胞来自小鼠肺和淋巴器官的亚群（内皮细胞）。 Core 将允许通过 RNA 测序对转录组反应进行细胞类型特异性评估，包括对于这一点，Core 将提供从分选细胞中自动分离 DNA/RNA 的功能。随后进行质量评估和RNA测序文库构建以及Core C测序。小鼠产生细胞类型或组织特异性 Cre 重组酶系以诱导组织特异性敲除 Core C 将对项目负责人提出的所有小鼠品系进行基因分型。维持野生型和基因工程小鼠的统一环境，遭受肺损伤并允许康复，这些工具为项目提供了独特的资源。调查人员，如果没有这个 PPG 的支持，这将很难重现。