Scientific Core

科学核心

• 批准号: 8289361
• 负责人: Jane B. Lian
• 金额: $ 9.3万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8289361
• 关键词: Adenoviruses; Animals; Antibodies; Breeding; Calvaria; Cell Culture Techniques; Cell Line; Cells; Computer software; Culture Media; Differentiation and Growth; Embryo; Ensure; Enzymes; Freezing; Gene Expression; Genotype; Histology; Image; Immunofluorescence Immunologic; In Situ; Knock-in Mouse; Maintenance; Marrow; Microscope; Microscopy; Monitor; Mus; Mutant Strains Mice; Nuclear Matrix-Associated Proteins; Osteoblasts; Paraffin Embedding; Phenotype; Plastic Embedding; Preparation; Procedures; Program Research Project Grants; Property; Quality Control; RNA; Rattus; Reagent; Runx2 protein; Serum; Services; Southern Blotting; Staining method; Stains; Tissues; Transgenes; Transgenic Organisms; bone cell; data management; fetal; genetic regulatory protein; mouse model; mutant; sterility testing; transcription factor

The Scientific Core has three major components: 1) Cell cultures, 2) Animals and 3) Microscopy. The Core will assist each of the projects by providing reagents and support services. These include characterized cells, breeding, genotyping, and phenotyping of mouse models, microscopy, in situ immunofluorescence imaging and histology. Scientific and technical support includes: 1. Routine isolation and maintenance of primary fetal rat calvarial-derived osteoblasts, marrow cells and calvarial osteoblasts from transgenic/knock-in mice as needed. 2. Maintenance of various osseous and non-osseous cell lines and cell lines with transgenes. Preparation and sterility testing of cell culture media, screeening of serum lots and enzyme reagents to ensure uniformity of cultures. 3. Monitoring growth and differentiation properties of cell lines for phenotype stability. 4. Preparation of adenovirus stocks of Runx2 proteins. 5. Maintenance of Runx2 knock-in/mutant mouse colonies as required for all three projects. This includes breeding, tagging, and identifying positive and negative mice by Southern blot or PCR analysis. 6. Routine preparation of embryos or tissues for phenotype characterization, which may include either whole embryo (3-Gal staining, frozen, plastic or paraffin embedding, sectioning and staining, and jnCT analyses. 7. Preparation of total cellular RNA and nuclear matrix proteins from tissues and cell lines. 8. Characterization and quality control of all antibodies used for in situ immunofluorescence. 9. Maintenance of all microscopes and software applications. 10. Data management of quantitative procedures to characterize distribution of wild type and mutant Runx2 transcription factors and their co-regulatory proteins.

科学核心具有三个主要组成部分：1）细胞培养物，2）动物和3）显微镜。核心 将通过提供试剂和支持服务来帮助每个项目。这些包括特征 小鼠模型的细胞，繁殖，基因分型和表型，显微镜，原位免疫荧光 成像和组织学。科学和技术支持包括： 1。定期隔离和维持原发性胎儿大鼠钙钙衍生的成骨细胞，骨髓细胞和 根据需要来自转基因/敲门小鼠的颅骨成骨细胞。 2。维持各种骨和非骨的细胞系以及具有转基因的细胞系。准备 以及细胞培养基的无菌测试，血清批次和酶试剂的筛选，以确保 文化的统一性。 3。监测细胞系的生长和分化特性以进行表型稳定性。 4。制备RUNX2蛋白的腺病毒库存。 5。根据所有三个项目所需的需要维护Runx2敲入/突变小鼠菌落。这包括 通过Southern印迹或PCR分析，繁殖，标记和识别阳性和负小鼠。 6。胚胎或组织的常规制备以进行表型表征，可能包括 整个胚胎（3-GAL染色，冷冻，塑料或石蜡嵌入，切片和染色，JNCT 分析。 7。从组织和细胞系中制备总细胞RNA和核基质蛋白。 8。用于原位免疫荧光的所有抗体的表征和质量控制。 9。维护所有显微镜和软件应用程序。 10。定量程序的数据管理，以表征野生型和突变体Runx2的分布 转录因子及其共调节蛋白。