Animal Core

动物核心

基本信息

批准号：
8561432
负责人：
Guido Erwin Michael Eibl
金额：
$ 25.83万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8561432
关键词：
Alleles Animal Euthanasia Animal Housing Animal Model Animals Area Back Biological Biometry Blood Breeding Cell Culture Techniques Comprehensive Cancer Center DNA Data Data Collection Development Diet Digestive System Disorders Drug or chemical Tissue Distribution Ensure Environment Evaluation Event Fatty acid glycerol esters Formalin Freezing Generations Genetic Genetic Recombination Genetically Engineered Mouse Genotype Guidelines Harvest Histology Human Human Resources Immunohistochemistry Individual Instruction Intervention Laboratories Laboratory Research Liquid substance Logistics Malignant neoplasm of pancreas Medical center Modeling Monitor Mus Mutant Strains Mice Mutate Nitrogen Organ Pancreas Pancreatic Adenocarcinoma Pancreatic Diseases Paraffin Pathology Phase Preparation Procedures Process Protein Analysis RNA analysis Randomized Research Personnel Resource Sharing Resources Sampling Services Tail Tissue Banking Tissue Banks Tissue Harvesting Tissues Transgenes Transgenic Mice Work animal care animal facility animal tissue cohort cost design embryonic stem cell meetings member mouse model offspring programs research study square foot tissue processing training project tumor

项目摘要

The Animal Core will provide the main animal model (conditional Kras[G12D] mouse model) to all four projects. Heterozygous LSL-Kras[G12D/+] mice will be crossed with heterozygous Pdx-1-Cre mice. Offspring will be genotyped for the presence of the mutated Kras allele and Cre transgene using PCR analysis of mouse tail DNA. Mutant mice {LSL-Kras[G12D/+];Pdx-1-Cre) will be randomly allocated to either the control diet (AIN-76A) or the high fat, high calorie diet (HFCD). Each of the experimental groups will then be allocated to the individual project-specific treatments and interventions. The Animal Core will conduct all aspects related to the Project-specific animal studies. Cohorts of mice will be sacrificed at 3, 6, 9, and 12 months or prematurely terminated according to the institutional guidelines. At sacrifice, tissues, including the entire pancreas, other organs and blood will be harvested and processed. The exact post-processing procedures depend on the individual Project requirements and needs. In general, half of the pancreas will be fixed in formalin and embedded in paraffin (for standard histology and immunohistochemistry), the other half frozen in liquid nitrogen (for RNA and protein analysis). A small piece of the pancreas will be used for genotyping (to detect the successful recombination event). Included in the Animal Core is the Pathology Sub-Core, which will provide the histo-pathological evaluation of all mouse tissues. Services that the Animal Core provides for each Project include: Obtaining and maintaining institutional approval, designing the animal studies in terms of logistics and statistical power calculations (together with Project Pi's and Biostatistics Sub-Core), maintaining animal strains and setting up breeding pairs, genotyping of all animals, randomization of animals to experimental groups, preparation and administration of experimental diets or interventions, daily monitoring and animal care, euthanasia of animals, harvest of tissues and blood at sacrifice, processing tissue, e.g. fixing in formalin and embedding in paraffin, and sectioning (together with the Pathology Sub-Core), histopathological evaluation of tissue sections (together with the Pathology Sub- Core), generation of cell cultures (PanlNs and murine pancreatic cancer), storage of raw data, central banking of tissues and biological samples, distribution of tissues to individual Projects, discussion of results with Project Leaders, training of Project investigators in animal procedures (if desired).

Animal Core 将为所有四个项目提供主要动物模型（条件 Kras[G12D] 小鼠模型）。杂合 LSL-Kras[G12D/+] 小鼠将与杂合 Pdx-1-Cre 小鼠杂交。将使用小鼠尾部 DNA 的 PCR 分析对后代进行基因分型，以确定是否存在突变的 Kras 等位基因和 Cre 转基因。突变小鼠{LSL-Kras[G12D/+]；Pdx-1-Cre)将被随机分配到对照饮食(AIN-76A)或高脂肪、高热量饮食(HFCD)。然后，每个实验组将被分配到具体项目的治疗和干预措施。动物核心将开展与项目特定动物研究相关的所有方面。根据机构指南，小鼠群体将在 3、6、9 和 12 个月时被处死或提前终止。处死时，将收集并处理组织，包括整个胰腺、其他器官和血液。确切的后处理程序取决于各个项目的要求和需求。一般来说，一半胰腺将固定在福尔马林中并包埋在石蜡中（用于标准组织学和免疫组织化学），另一半冷冻在液氮中（用于 RNA 和蛋白质分析）。一小块胰腺将用于基因分型（以检测成功的重组事件）。动物核心中包括病理学子核心，它将提供所有小鼠组织的组织病理学评估。动物核心为每个项目提供的服务包括：获得并维持机构批准、根据物流和统计功效计算设计动物研究（连同 Project Pi 和生物统计子核心）、维护动物品系并建立育种对，所有动物的基因分型、将动物随机分配到实验组、实验饮食或干预措施的准备和管理、日常监测和动物护理、动物安乐死、处死时收获组织和血液、处理组织，例如组织。在福尔马林中固定并在石蜡中包埋，切片（与病理学子核心一起），组织切片的组织病理学评估（与病理学子核心一起），细胞培养物的产生（PanlNs和鼠胰腺癌），原始细胞的储存数据、组织和生物样本的中央银行、向各个项目分配组织、与项目负责人讨论结果、对项目研究人员进行动物程序培训（如果需要）。