Cell Biology and Multigene Transcriptional Profiling Core

细胞生物学和多基因转录分析核心

• 批准号: 8378445
• 负责人: John W LAWLER
• 金额: $ 18.55万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-13 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8378445
• 关键词: Antibodies; Apoptosis; Biological Preservation; Breast Cancer Cell; Cancer cell line; Cell Culture Techniques; Cell Cycle; Cell Line; Cells; Cellular biology; Characteristics; Colon Carcinoma; Complementary DNA; Consult; Cultured Cells; Data; Data Set; Dermal; Endothelial Cells; Ensure; Gene Expression; Gene Expression Profiling; Genes; Growth; Growth Factor; Heart; Human; Individual; Lead; Lewis Lung Carcinoma; Libraries; Lung; Maintenance; Malignant Epithelial Cell; Measurement; Measures; Messenger RNA; Molecular Profiling; Monitor; Mus; Names; Phenotype; Play; Process; Quality Control; Rattus; Reagent; Regulation; Research Personnel; Sampling; Serum; Statistical Data Interpretation; Techniques; Testing; Time; Umbilical vein; Validation; angiogenesis; base; design; dimer; fetal; glioma cell line; meetings; novel marker; programs; repository; spelling; tissue culture

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.

细胞生物学和基因分析核心将提供标准化，同质和良好的细胞 人类和老鼠的文化来源于各个项目的研究者。此外，这个核心将 提供多基因的转录分析（MGTP），以表征培养和组织中细胞的表征。 核心将产生人类皮肤内皮细胞（HDMEC），小鼠心脏和肺内皮细胞， ymphatic内皮细胞，以及人脐静脉内皮细胞（HUVEC）的大规模扩增。 核心已经开发了一系列用于内皮细胞标记的抗体，这将用于监测 保存培养物中表型的保存，并将充当由细胞系的存储库 OROGRAM。细胞生物学和基因分析核心也将负责质量控制测试 某些关键试剂（例如胎儿血清，生长因素，媒体）以及这些试剂的大量购买 所有项目的使用。此功能确保培养细胞的一致生长和维持条件 从核心在核心内部的隔离和传播中，通过他们对各个人的操纵 项目。该核心还将培养项目中将使用的各种癌细胞系。 通过MGTP（多基因的转录分析），此核心也可以向研究人员提供 单个项目，用于准确定量基因表达的设施。 MGTP是一种基因表达 基于定量实时PCR的分析技术使用设计用于均匀扩增的引物 用于多基因面板的高实验吞吐量的效率和主cDNA模板。 MGTP是 能够准确且可重复测量每个细胞表达的mRNA拷贝数。我们的底漆 库当前包括1000多种不同的人和鼠标基因，它们在 血管生成，细胞周期和凋亡，我们将继续增加数百个文库 每年基因。核心将在基因分析数据的统计分析中提供专业知识。 MGTP 还将用于表征由核心制备的HDMEC，以确保批次对路线一致性 并开发新标记。