Cell Biology and Multigene Transcriptional Profiling Core

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The Cell Biology and Gene Profiling Core will provide standardized, homogenous and well-characterized cell cultures of human and mouse origin to the investigators of the individual projects. In addition, this Core will provide multi-gene transcriptional profiling (MGTP) for the characterization of cells in culture and tissues. The core will produce human dermal endothelial cells (HDMEC), mouse heart and lung endothelial cells, ymphatic endothelial cells, and large-scale amplification of human umbilical vein endothelial cells (HUVEC). The core has developed a panel of antibodies for endothelial cell markersthat will be used to monitor preservation of phenotype in culture and will function as a repository for cell lines that are generated by the orogram. The Cell Biology and Gene Profiling Core will also be responsible for quality control testing of certain key reagents (e.g. fetal serum, growth factors, media) and the bulk purchase of these reagents for the use of all projects. This function ensures consistent growth and maintenance conditions for cultured cells from their isolation and propagation within the core through their manipulation in the various individual projects. This core will also culture the various cancer cell lines that will be used within the project. Through MGTP (multi-gene transcriptional profiling), this core also makes available to the investigators of the individual projects a facility for accurate quantification of gene expression. MGTP is a gene expression profiling technique based on quantitative real-time PCR using primers designed for uniform amplification efficiencies and a master cDNA template for high experimental throughput for multi-gene panels. MGTP is capable of accurately and reproducibly measuring mRNA copy numbers expressed per cell. Our primer library currently includes over 1000 different human and mouse genes that play critical functions in angiogenesis, the cell cycle, and apoptosis, and we will continue to grow the library by several hundred genes per year. The Core will provide expertise in the statistical analysis of the gene profiling data. MGTP will also be used to characterize the HDMEC that are prepared by the core to assure lot-to-lot consistency and to develop new markers.