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.

动物核心将为所有四个项目提供主要动物模型（有条件的Kras'^^^^小鼠模型）。杂合LSL-KRAS'^^^*小鼠将与杂合PDX-1-CRE小鼠交叉。后代将是使用小鼠尾巴的PCR分析，用于突变的KRAS等位基因和CRE转基因的基因分型脱氧核糖核酸。突变小鼠{lsl-kras¿ 或高脂肪高热量饮食（HFCD）。然后将每个实验组分配给各个项目特定的治疗和INTEN/ENFIONS。动物核心将进行与特定于项目的动物研究。小鼠队列将在3、6、9和12个月或12个月或根据机构指南过早终止。在牺牲中，组织，包括整个胰腺，其他器官和血液将被收获和加工。确切的后处理程序取决于单个项目的要求和需求。通常，一半的胰腺将被固定在福尔马林和嵌入石蜡（用于标准的组织学和免疫组织化学）中，另一半冷冻在液氮中（用于RNA和蛋白质分析）。一小部分胰腺将用于基因分型（检测成功的重组事件）。动物核心包括病理亚核，这将提供所有小鼠触发的组织病理评估。动物核心的服务提供每个项目包括：获得和维持机构批准，设计动物根据逻辑和统计功率计算的研究（与项目PI和生物统计学一起亚核），维持动物菌株和建立繁殖对，所有动物的基因分型，动物向实验组随机化，实验饮食的准备和给药或 Inten/Entions，日常监测和动物护理，动物安乐死，组织的收获和血液牺牲，加工组织，例如固定福尔马林并嵌入石蜡，并与病理亚核，组织切片的组织病理学评估（以及病理亚核心），生成细胞培养物（Panlns和鼠胰腺癌），原始数据的存储，中央分支和生物样品的押金，分配给单个项目的兄弟会，讨论结果在项目负责人的情况下，对项目调查人员进行动物程序的培训（如果需要）。相关性（请参阅说明）：动物核心的对象fives是通过协调来最大程度地减少重复的努力并降低成本不同项目研究人员进行的实验，以便动物和组织可以由调查人员，如有可能。通过集中化，动物研究所需的资源将是非常有效地利用（与单独进行动物研究的单个项目相比）。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。