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).
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.
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