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. !
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