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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（条件性Kras[G12 D]小鼠模型）。杂合LSL-Kras[G12 D/+]小鼠将与杂合Pdx-1-Cre小鼠杂交。使用小鼠尾部DNA的PCR分析，对后代进行突变的Kras等位基因和Cre转基因的基因分型。将突变小鼠（LSL-Kras[G12 D/+];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）。然后，每个实验组将被分配到个别项目特定的治疗和干预措施。动物中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，在3、6、9和12个月时处死小鼠队列或提前终止。在处死时，将采集和处理组织，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，一半的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，其将提供所有小鼠组织的组织病理学评价。Animal Core为每个项目提供的服务包括：获得并保持机构批准，在后勤和统计功效计算方面设计动物研究（与Project Pi和生物统计子核心一起），保持动物品系并建立繁殖对，对所有动物进行基因分型，将动物随机分配到实验组，制备和给予实验饮食或干预措施，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，例如福尔马林固定和石蜡包埋，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），细胞培养物的生成（PanIN和鼠胰腺癌），原始数据的储存，组织和生物样本的中央库，组织分配到各个项目，与项目负责人讨论结果，对项目研究者进行动物程序培训（如需要）。