All animal experiments were approved by the Memorial Sloan Kettering Cancer Center (MSKCC) Animal Care and Use Committee. Female nude mice (6 weeks old) were obtained from ENVIGO, female B6 129F1 (6 weeks old) were obtained from TACONIC, female B6 FVBF1/J (6 weeks) and female NSG mice (6 weeks) were obtained from JACKSON LABORATORY, and housed in accredited facilities under pathogen-free conditions. Additional information on experimental methods in next section.
Female nude mice
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Female nude mice are genetically engineered laboratory animals that lack a functional adaptive immune system, including the thymus and lymphoid tissues. These mice are commonly used in biomedical research to study disease processes and evaluate therapeutic interventions.
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10 protocols using female nude mice
1
Nude Mice Animal Experiments
2
Mitigating Breast Cancer Tumor Growth
3
In Vivo Metastasis Induction
4
Tumor Xenograft Model for Vitamin B12 Imaging
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All of the animal
handling and manipulations were conducted in accordance with the guidelines
set by WSU Animal Care and Use Committee (IACUC). For imaging and
in vivo uptake experiments, female nude mice (Envigo) were kept under
Cbl-deficient diet (Teklad Cbl-free custom diet, Envigo) for 3 weeks.
Cells were subcutaneously implanted on the shoulder with MDA-MB-453
cancer cells (5 × 106 cells/mouse) after 2 weeks of
Cbl-free diet. Cells were injected in 1:1 media/matrigel (Corning
LLC) at a volume of 200 μL. The tumor volume until was calculated
using the formula length × width2 × 0.52. Mice
with tumors of 100–200 mm3 dimensions were used
for imaging experiments.
handling and manipulations were conducted in accordance with the guidelines
set by WSU Animal Care and Use Committee (IACUC). For imaging and
in vivo uptake experiments, female nude mice (Envigo) were kept under
Cbl-deficient diet (Teklad Cbl-free custom diet, Envigo) for 3 weeks.
Cells were subcutaneously implanted on the shoulder with MDA-MB-453
cancer cells (5 × 106 cells/mouse) after 2 weeks of
Cbl-free diet. Cells were injected in 1:1 media/matrigel (Corning
LLC) at a volume of 200 μL. The tumor volume until was calculated
using the formula length × width2 × 0.52. Mice
with tumors of 100–200 mm3 dimensions were used
for imaging experiments.
5
Dexamethasone Modulates Tumor Growth
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Female nude mice at 4–5 weeks old were purchased from Envigo. KMM cells were injected into both flanks of 32 mice at 107 cells per site. Each mouse generates two tumors. In the stage I experiment, mice were randomly split into two groups to receive either 20 mg/kg dexamethasone (Dex, n = 16) or vehicle treatment (ethanol in phosphate buffer saline, Vehicle, n = 16) starting at day 17 post-inoculation by intraperitoneal injection for 5 days per week (day 17–50). In stage II experiment, the vehicle group was evenly divided into two subgroups at day 56, and one subgroup was treated with dexamethasone (Vehicle-Dex, n = 8), while the second subgroup remained untreated (Vehicle-Vehicle, n = 8). Meanwhile, the dexamethasone group was evenly divided into two subgroups, and one subgroup remained treated with dexamethasone (Dex-Dex, n = 8), while the second subgroup was no longer treated (Dex-Vehicle, n = 8). Treatment was given 5 days per week. The initial vehicle group was terminated at day 77, while the initial dexamethasone group was terminated at day 140, when the volumes of some tumors reached 1.5 cm3. Tumor incidence and growth were analyzed as previously described (41 (link)). The body weights of the mice and tumor volumes were measured every 3 days.
6
Conditional Deletion of Mdm2 and p53 in Mice
7
In Vivo Anti-Tumor Studies in Nude Mice
8
Angiogenesis Inhibition in Xenograft Models
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DU145 cells were injected intradermally into male nude mice (Envigo), and MDA-MB-231 cells were injected intradermally into female nude mice (Envigo). Cells were injected at four sites on the ventral side of the animal, at 105 cells in 10 μL of PBS. Three mice were used per treatment group. Mice either received cells that were exposed to BMS-777607 for 24 hours prior to injection, or the mice received the drug orally every day for the duration of the experiment. Three days after tumor cell injection, mice were euthanized, skin flaps were removed, and the number of blood vessels growing into the tumor nodule was quantified visually on a stereoscope.
9
Nude Mice Animal Experiments
10
Xenograft Tumor Formation in Mice
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Female nude mice were purchased from Envigo (Horst, the Netherlands). After 1 week of adaptation to the husbandry, the mice entered the experiment at approximately 7e8 weeks of age. The experiments were approved by the Cantonal Veterinary Office of Zurich (license ZH175/18) and performed according to Swiss laws and regulations. Briefly, as previously described (Freiberger et al., 2015; Hofbauer et al., 2014) , 100 ml of 1 Â 10 6 wild-type or transfected SSC cells or 4 Â 10 6 wild-type or transfected HaCaT cells were subcutaneously injected into the flank of athymic nude mice. A total of 10 mice per group were used in all experiments. Mice weight was monitored weekly, and the tumor size was measured thrice a week using a caliper. The tumor volume was calculated using the formula V ¼ l 1 Âl 2 /2, l 1 and l 2 being the long and short diameters, respectively. Individual mice were euthanatized if tumor volume exceeded 1.5 cm 3 , if tumors ulcerated, upon severe weight loss, or upon severely reduced health conditions. On termination of experiments, tumors were excised and bisected for protein extraction and histology. Lungs were excised and fixed in phosphate-buffered paraformaldehyde for further analyses.
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