Animal Core

动物核心

基本信息

批准号：
8373952
负责人：
Guido Erwin Michael Eibl
金额：
$ 27.01万
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：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8373952
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 Recombination Genetically Engineered Mouse Genotype Guidelines Harvest Histology Human Human Resources Immunohistochemistry Individual Instruction Intervention Laboratories Laboratory Research Liquid substance Logistics Malignant Neoplasms 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'^^^^ mouse model) to all four projects. Heterozygous LSL-Kras'^^^¿^* 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¿^^¿'^;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 inten/enfions. 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 evaluafion of all mouse fissues. Services that the Animal Core provides for each Project include: Obtaining and maintaining institutional approval, designing the animal studies in terms of logisfics 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 inten/entions, 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 secfioning (together with the Pathology Sub-Core), histopathological evaluation of tissue sections (together with the Pathology Sub- Core), generation of cell cultures (PanlNs and murine pancreafic cancer), storage of raw data, central banking of fissues and biological samples, distribufion of fissues to individual Projects, discussion of results with Project Leaders, training of Project investigators in animal procedures (if desired). RELEVANCE (See instructions): The Animal Core's objecfives are to minimize duplication of effort and reduce costs by coordinating the experiments performed by different Project Investigators so that animals and tissues may be shared by the investigators, where possible. Through the centralization, resources needed for the animal studies will be utilized very efficiently (as compared to the individual Projects performing their animal studies separately). Providing centralized animal services will ensure consistency in animal handling and data collection.

Animal Core 将为所有四个项目提供主要动物模型（条件 Kras'^^^^ 小鼠模型）。杂合 LSL-Kras'^^^¿ ^* 小鼠将与杂合 Pdx-1-Cre 小鼠杂交，产生后代。使用小鼠尾部的 PCR 分析对突变的 Kras 等位基因和 Cre 转基因的存在进行基因分型 DNA突变小鼠{LSL-Kras¿ ^^¿ '^;Pdx-1-Cre) 将被随机分配到对照饮食 (AIN-76A) 然后将每个实验组分配到高脂肪、高热量饮食（HFCD）。个别项目特定的治疗和意图/enfion 将进行与相关的所有方面。特定项目的动物研究将在 3、6、9 和 12 个月或根据机构指南提前终止处死组织，包括整个组织。胰腺、其他器官和血液将被采集并进行精确的后处理程序。取决于个人项目的要求和需要，一般来说，一半的胰腺会被固定。福尔马林并包埋在石蜡中（用于标准组织学和免疫组织化学），另一半冷冻液氮中（用于 RNA 和蛋白质分析）。（检测成功的重组事件）动物核心中包含病理学子核心，这将提供动物核心提供的所有小鼠组织服务的组织病理学评估。为每个项目提供的规定包括：获得并维持机构批准、设计动物逻辑学和统计功效计算方面的研究（与 Project Pi's 和 Biostatistics 一起）子核心），维护动物品系并建立繁殖对，对所有动物进行基因分型，将动物随机分配到实验组、实验饮食的制备和管理或意图/意图、日常监测和动物护理、动物安乐死、采集组织和血液牺牲，处理组织，例如在福尔马林中固定并包埋在石蜡中，以及切片（连同病理学子核心），组织切片的组织病理学评估（与病理学子核心一起）核心），细胞培养物的产生（PanlNs和小鼠胰腺癌），原始数据的存储，中央组织和生物样本的银行、将组织分配给各个项目、结果讨论与项目负责人一起，对项目调查员进行动物程序方面的培训（如果需要）。相关性（参见说明）： Animal Core 的目标是通过协调各个部门来最大程度地减少重复工作并降低成本由不同的项目研究人员进行的实验，以便动物和组织可以由不同的项目研究人员共享在可能的情况下，通过集中化，将获得动物研究所需的资源。利用率非常高（与单独进行动物研究的各个项目相比）。提供集中的动物服务将确保动物处理和数据收集的一致性。