Cell Biology and Multigene Transcriptional Profiling Core

细胞生物学和基因图谱核心将提供标准化、同质和特征良好的细胞 向个别项目的研究人员提供人类和鼠类来源的培养。此外，这一核心将 提供多基因转录图谱(MGTP)，用于鉴定培养和组织中的细胞。 核心将产生人真皮内皮细胞(HDMEC)、小鼠心脏和肺内皮细胞， 淋巴管内皮细胞和人脐静脉内皮细胞的大规模扩增。 CORE已经开发了一组针对内皮细胞标志物的抗体，将用于监测 在培养中保存表型，并将作为由 字形。细胞生物学和基因图谱核心还将负责质量控制测试 某些关键试剂(如胎儿血清、生长因子、培养液)和批量购买这些试剂用于 所有项目的使用。此功能可确保培养细胞的生长和维持条件一致 从它们在核心内的隔离和传播通过它们在不同个体中的操纵 项目。这个核心还将培养项目中将使用的各种癌细胞株。 通过MGTP(多基因转录图谱)，这一核心也使 个体计划为基因表达的准确量化提供了一种工具。MGTP是一种基因表达 基于均匀扩增设计的实时荧光定量聚合酶链式反应技术 效率和用于多基因面板的高实验吞吐量的主cdna模板。MGTP是 能够准确和重复性地测量每个细胞表达的mRNA拷贝数。我们的初级读物 文库目前包括1000多个不同的人类和老鼠基因，这些基因在 血管生成、细胞周期和细胞凋亡，我们将继续扩大几百个文库 每年的基因数量。该中心将提供基因图谱数据的统计分析方面的专业知识。MGTP 还将用于表征由核心准备的HDMEC，以确保批次到批次的一致性 并开发新的标记。

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.