Scientific Core

科学核心有三个主要组成部分：1)细胞培养，2)动物和3)显微镜。《核心》 将通过提供试剂和支持服务来协助每个项目。其中包括特色化 细胞，小鼠模型的育种，基因分型和表型，显微镜，原位免疫荧光 影像和组织学。科学技术支持包括： 1.原代胎鼠颅骨来源成骨细胞、骨髓细胞的常规分离和维持 根据需要从转基因/敲入小鼠获得颅骨成骨细胞。 2.各种骨和非骨细胞系及转基因细胞系的维持。制备 以及细胞培养液的无菌测试，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确保表型稳定性。 4.Runx2蛋白重组腺病毒的制备。 5.维护所有三个项目所需的Runx2基因敲入/突变小鼠群体。这包括 通过Southern杂交或聚合酶链式反应分析，培育、标记和鉴定阳性和阴性小鼠。 6.用于表型鉴定的胚胎或组织的常规准备，可包括 完整胚胎(3-Gal染色、冷冻、塑料或石蜡包埋、切片和染色以及联合胚胎移植) 分析。 7.从组织和细胞系中制备细胞总RNA和核基质蛋白。 8.用于原位免疫荧光的所有抗体的特性和质量控制。 9.维护所有显微镜和软件应用程序。 10.描述野生型和突变型Runx2分布的定量程序的数据管理 转录因子及其共调控蛋白。

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.