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，同质和良好的表征细胞 人和小鼠来源的培养物提供给各个项目的研究人员。此外，该核心将 提供多基因转录谱分析（MGTP），用于表征培养物和组织中的细胞。 该核心将产生人真皮内皮细胞（HDMEC）、小鼠心脏和肺内皮细胞， 淋巴管内皮细胞和大规模扩增人脐静脉内皮细胞（HUVEC）。 该中心已经开发了一组针对内皮细胞标记的抗体， 在培养物中保存表型，并将作为细胞系的储存库， 地形图细胞生物学和基因分析核心还将负责质量控制测试， 某些关键试剂（例如，胎儿血清、生长因子、培养基）和批量购买这些试剂， 所有项目的使用。该功能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，到它们在各个个体中的操纵， 项目该核心还将培养将在项目中使用的各种癌细胞系。 通过MGTP（多基因转录谱），该核心也可供研究人员使用。 个人项目的设施，精确定量的基因表达。MGTP是一种基因表达 基于使用设计用于均匀扩增的引物的定量实时PCR的谱分析技术 高效率和主cDNA模板，用于多基因组的高实验通量。MGTP是 能够准确且可重复地测定每个细胞表达的mRNA拷贝数。我们米特尔 文库目前包括1000多个不同的人类和小鼠基因，这些基因在人类和小鼠的基因组中发挥关键作用。 血管生成，细胞周期和凋亡，我们将继续增加数百个库 基因每年核心将提供基因图谱数据统计分析方面的专门知识。MGTP 还将用于表征核心制备的HDMEC，以确保批次间的一致性 并开发新的标记。