Animal Core

动物中心将为所有四个项目提供主要动物模型（条件性Kras小鼠模型）。 将杂合LSL-Kras-1-Cre小鼠与杂合Pdx-1-Cre小鼠杂交。后代将是 使用PCR分析小鼠尾部的突变Kras等位基因和Cre转基因的存在进行基因分型 DNA.将突变小鼠（LSL-Kras;Pdx-1-Cre）随机分配至对照饮食（AIN-76 A） 或高脂肪高热量饮食（HFCD）。每个实验组将被分配到 个别项目特定的治疗和干预。动物中心将进行与以下方面有关的所有方面： 特定项目的动物研究。将在3、6、9和12个月时处死小鼠组，或 根据机构指南提前终止。在处死时，组织，包括整个 胰腺、其他器官和血液将被收获和处理。准确的后处理程序 取决于各个项目的要求和需求。一般来说，一半的胰腺将被固定在 福尔马林和石蜡包埋（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 (to检测成功的重组事件）。动物核心包括病理学子核心， 这将提供所有小鼠骨折的组织病理学评价。Animal Core提供的服务 每个项目的规定包括：获得和保持机构批准，设计动物 逻辑学和统计功效计算方面的研究（与Project Pi和生物统计学一起 子核心），保持动物品系和建立育种对，对所有动物进行基因分型， 将动物随机分配至实验组，制备和给予实验饲料，或 实验、日常监测和动物护理、动物安乐死、组织和血液采集 处死，处理组织，例如在福尔马林中固定并在石蜡中包埋，以及切片（连同 病理学子核心）、组织切片的组织病理学评价（连同病理学子核心） 核心）、细胞培养物（PanIN和鼠胰腺癌）的产生、原始数据的存储、中心数据库的建立、细胞培养物（PanIN和鼠胰腺癌）的 储存生物样品和生物样品，将生物样品分配给各个项目，讨论结果 与项目负责人一起，对项目研究者进行动物程序培训（如需要）。 相关性（参见说明）： 动物核心的目标是通过协调 由不同的项目研究者进行的实验，以便动物和组织可以由 调查人员，如果可能的话。通过集中化，动物研究所需的资源将被 非常有效地利用（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。

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.