Core B: The Cell Culture Core

项目概要：核心B 细胞培养核心，指定为核心B，是一个集中的设施，提供原代小鼠胚胎 将从野生型和遗传修饰的小鼠分离的成纤维细胞、内皮细胞和上皮细胞分别接种到每种细胞中。 五个项目。核心B还维持PPG的次级肺细胞系，并将产生基因组编辑的 细胞核心B将使用先进的基因组编辑技术来产生表达mEmerald-vimentin的细胞， 在其内源性启动子的控制下，以及基于CRISPR/Cas9的基因组编辑， 缺乏波形蛋白的细胞。核心人员在原代小鼠隔离方面具有丰富的经验 胚胎成纤维细胞、组织特异性成纤维细胞、内皮细胞和来自小鼠组织（例如肺）的上皮细胞 和肝）以及一般的细胞培养技术。细胞培养设施的集中化将确保 五个项目均会持续供应优质电池。巩固的细胞 培养设备提供了分离和培养原代和次级细胞的经济手段。这 转化为降低的总成本（即，人员、试剂和动物），更重要的是， 用于研究的细胞的质量。核心还将为研究人员提供几种鼠类品系， 包括vimentinfl/fl小鼠，这将使研究人员能够检查基因或途径的细胞特异性作用。 使用表达Cre重组酶的小鼠或使用他莫昔芬诱导系统。许多菌株将被用于 在一个以上的项目中，强调本计划项目可实现的协同作用和规模经济。 这些动物将用于生成项目研究者概述的研究所需的细胞和组织。 此外，Core B将进行有限的RNA分析，以评估波形蛋白中的特异性基因表达 使用西北大学下一代测序（NGS）核心， 设施。 !

Project Summary: Core B The Cell Culture Core, designated as Core B, is a centralized facility that provides primary mouse embryonic fibroblasts, endothelial and epithelial cells isolated from wild-type and genetically modified mice to each of the five projects. Core B also maintains secondary lung cell lines for the PPG and will generate genome-edited cells. Core B will use advanced genome-editing technology to produce cells expressing mEmerald-vimentin, both under the control of its endogenous promoter, as well CRISPR/Cas9-based genome editing to produce cells lacking vimentin protein. The Core personnel have extensive experience in the isolation of primary mouse embryonic fibroblasts, tissue-specific fibroblasts, endothelial, and epithelial cells from mouse tissue (e.g. lung and liver), as well as general cell culture techniques. Centralization of the cell culture facilities will ensure that continuous supply of high-quality cells will be available to each of the five projects. The consolidated cell culture facility provides an economical means of isolating and culturing primary and secondary cells. This translates into reduced overall costs (i.e., personnel, reagents, and animals) and more importantly maintains the quality of the cells used in research. The Core will also provide investigators several murine strains, including vimentinfl/fl mice, which will allow investigators to examine cell-specific roles of genes or pathways using mice expressing Cre recombinase or using tamoxifen-inducible systems. Many of the strains will be used in more than one project, highlighting the synergy and economies of scale achievable by this Program Project. These animals will be used to generate cells and tissues for investigations outlined by the project investigators. In addition, Core B will conduct limited RNA analysis to assess the specific gene expression in vimentin expressing and vimentin-null cells using the Northwestern University Next-Generation Sequencing (NGS) Core Facility. !