Animal Core

动物核心将为所有四个项目提供主要的动物模型(条件性Kras[G12D]小鼠模型)。杂合子LSL-Kras[G12D/]小鼠将与杂合子PDX-1-Cre小鼠杂交。后代将通过对小鼠尾部DNA的聚合酶链式反应分析，对是否存在突变的Kras等位基因和Cre转基因进行基因分型。突变小鼠(LSL-Kras[G12D/]；PDX-1-CRE)将被随机分配到对照组(AIN-76A)或高脂肪、高热量饮食(HFCD)。然后，每个试验组将被分配到个别项目特定的治疗和干预措施。动物核心将进行与该项目特定动物研究相关的所有方面。根据机构的指导方针，小鼠队列将在3、6、9和12个月时被处死或提前终止。在牺牲时，组织，包括整个胰腺，其他器官和血液将被采集和处理。确切的后处理程序取决于个别项目的要求和需要。一般来说，一半的胰腺将固定在福尔马林中并包埋在石蜡中(用于标准的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)。动物核心包括病理学分核心，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：获得和维持机构批准，设计后勤和统计权力计算方面的动物研究(与PI项目和生物统计分核心一起)，维持动物品系并建立繁殖对，对所有动物进行基因分型，将动物随机分成试验组，准备和管理实验饲料或干预措施，日常监测和动物护理，动物的安乐死，牺牲时采集组织和血液，处理组织，例如福尔马林固定和石蜡包埋，以及切片(与病理分核心一起)，组织切片(连同病理学分核心)的组织病理学评估、细胞培养(Panlns和小鼠胰腺癌)的生成、原始数据的存储、组织和生物样本的中央银行、将组织分配给各个项目、与项目负责人讨论结果、对项目调查人员进行动物程序方面的培训(如果需要)。

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).