Female athymic nude mice (nu/nu, 4–6weeks old; Jackson Laboratories, USA) were randomized into 4 groups (n = 7). OVCAR5 SCG control-transfected and PLA2G3 KO cells (5 × 106 cells; 2 groups each) were injected intraperitoneally (i.p). Seven days following injection, one group each from the SCG and KO cohorts was treated with CBP (50 mg/kg) once in a week for 4 consecutive weeks. All mice euthanized on day30 and tumor weight was determined. Tumors were preserved in formalin for IHC and fresh frozen for protein analysis. The experiments were carried out under the approved protocol and guidelines of Mayo Clinic Animal Care and Use Committee, MN.
Female athymic nude mice nu nu
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Female athymic nude mice (nu/nu) are a strain of mice that lack a functional thymus gland, resulting in a deficiency of T cells. These mice are commonly used in biomedical research, particularly in studies involving immune function and the development of xenografts or tumor models.
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7 protocols using female athymic nude mice nu nu
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Ovarian cancer xenograft model
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In Vivo Xenograft Studies of OVCAR5 and ID8F3 Cancers
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In vivo studies were performed in the OVCAR5 xenograft model using female athymic nude mice (nu/nu, 4–6 weeks old; Jackson Laboratory, ME) and the ID8F3 syngeneic xenograft model using female C57BL/6 mice [37 (link)]. Details of the treatment modalities and assessment are discussed in the Supplementary section . Animal experiments were carried out under the approved protocols and guidelines of the Mayo Clinic Animal Care and Use Committee.
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Evaluating PLA2G3 KO in OVCAR5 Xenografts
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Female athymic nude mice (nu/nu, 4-6weeks old; Jackson Laboratories, USA) were randomized into 4 groups (n = 7). OVCAR5 SCG control-transfected and PLA2G3 KO cells (5×10 6 cells; 2 groups each) were injected intraperitoneally (i.p). Seven days following injection, one group each from the SCG and KO cohorts was treated with CBP (50 mg/kg) once in a week for 4 consecutive weeks. All mice euthanized on day30 and tumor weight was determined. Tumors were preserved in formalin for IHC and fresh frozen for protein analysis. The experiments were carried out under the approved protocol and guidelines of Mayo Clinic Animal Care and Use Committee, MN.
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Xenograft Models of Solid Tumors
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BALB/c female nude mice (obtained from Charles River, 7-8 weeks old) received unilateral subcutaneous injections of 200 μL of MDA-MB-468 cells (8 × 106 cells) suspended in phosphate buffered saline (PBS). For IGROV1 xenograft experiment, CD-1 female nude mice (obtained from Charles River, 8-9 weeks old) received unilateral subcutaneous injections of 100 μL of cells (2 × 106 cells) suspended in PBS. Once the tumors were palpable, subcutaneous growth was measured twice a week by caliper. Tumor volume was estimated using the following formula: length∗width2/2.
For HCT116 xenograft experiments, athymic female nude (nu/nu) mice (obtained from The Jackson Laboratory, 7-8 weeks old) received unilateral subcutaneous injections of 100 μL of HCT116 cells (2 × 106 cells) suspended in phosphate buffered saline (PBS). Once the different experimental groups reached an average tumor volume of approximatively 100 mm3, mice were treated either with vehicle (25% (w/v) hydroxypropyl- b-cyclodextrin in 10 mmol/L citrate, pH 2) or 200 mg/kg CB-839 (obtained from Calithera Bioscience) prepared in vehicle twice daily by oral gavage for 14 days. Subcutaneous growth was measured twice a week by caliper and tumor volume was calculated using the following formula: length∗width2/2. 4 hr after the last oral gavage, tumors were harvested and snap frozen for further analysis.
For HCT116 xenograft experiments, athymic female nude (nu/nu) mice (obtained from The Jackson Laboratory, 7-8 weeks old) received unilateral subcutaneous injections of 100 μL of HCT116 cells (2 × 106 cells) suspended in phosphate buffered saline (PBS). Once the different experimental groups reached an average tumor volume of approximatively 100 mm3, mice were treated either with vehicle (25% (w/v) hydroxypropyl- b-cyclodextrin in 10 mmol/L citrate, pH 2) or 200 mg/kg CB-839 (obtained from Calithera Bioscience) prepared in vehicle twice daily by oral gavage for 14 days. Subcutaneous growth was measured twice a week by caliper and tumor volume was calculated using the following formula: length∗width2/2. 4 hr after the last oral gavage, tumors were harvested and snap frozen for further analysis.
5
Mouse Xenograft Experiment Protocol
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All animal studies were conducted in compliance with UK Home Office approved project license and in accordance with institutional welfare guidelines. For xenograft experiments, BALB/c female nude mice (obtained from Charles River, 7-8 weeks old), CD-1 female nude mice (obtained from Charles River, 8-9 weeks old) and athymic female nude (nu/nu) mice (obtained from The Jackson Laboratory, 7-8 weeks old) were used. Mice were housed 5 per cage in a constant temperature (19-23°C) and humidity (55% ± 10%) animal room, with a 12-hour light/dark cycle (lights on at 7:00 am) and were allowed access to food and water ad libitum. Mice were allowed to acclimatize for one week prior to the experiment and were randomly assigned to experimental groups.
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Nude Mice Housing and Care
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Female athymic nude (Nu/Nu) mice were obtained from The Jackson Laboratory (Bar Harbor, ME) at 6 to 8 weeks of age. The mice were housed in a virus-free, indoor, light- and temperature controlled barrier environment, and were provided ad libitum access to food and water. All procedures with mice were in strict accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and approved by the Florida A & M University Animal Care and Use Committee.
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Cholesterol-based Liposomal 5-FU Delivery
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