Female nu nu athymic mice, by Inotiv - Product details

1

Xenograft Study of ARRB2 Overexpression

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All studies on mice were conducted using a protocol approved by the Augusta University Institutional Animal Care and Use Committee (IACUC). 1 × 10⁶ HT1376-EV or HT1376 ARRB2-OE cells were subcutaneously injected into eight week old athymic nu/nu female mice (Envigo Inc, Harlan, IN). We monitored tumor growth, and response to Gem therapy weekly. Tumor volume was measured three times weekly with a handheld manual caliper (Tumorimeter and RECIST Caliper [Cancer Technologies Inc.] (30 (link)). Gem (25 mg/kg bodyweight) was administered intraperitoneally, twice per week. Treatment was initiated after tumors reached an average volume of 100 mm³. Animal weight was measured weekly.

Kallifatidis G., Smith D.K., Morera D.S., Gao J., Hennig M.J., Hoy J.J., Pearce R.F., Dabke I.R., Li J., Merseburger A.S., Kuczyk M.A., Lokeshwar V.B, & Lokeshwar B.L. (2019). β-Arrestins regulate stem cell-like phenotype and response to chemotherapy in bladder cancer.. Molecular cancer therapeutics, 18(4), 801-811.

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2

Athymic Mouse Xenograft Model

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Athymic-nu/nu female mice aged 4 to 6 weeks were purchased from Envigo RMS Inc. (Indianapolis, IN). Mice were housed in the Animal Care Facility at the University of Iowa and all procedures were approved by the University of Iowa Institutional Animal Care and Use Committee and conformed to NIH guidelines. Three million FaDu tumor cells were injected subcutaneously into the right upper leg as described elsewhere (29 (link)).

Ma D.C., Anderson C.M., Rodman S.N., Buranasudja V., McCormick M.L., Davis A., Loth E., Bodeker K.L., Ahmann L., Parkhurst J.R., Sun W., Follmer K.R., Simons A.L., Buatt J.M., Spitz D.R., Fath M.A, & Allen B.G. (2021). Ketogenic Diet with Concurrent Chemoradiation in Head and Neck Squamous Cell Carcinoma: Preclinical and Phase 1 Trial Results. Radiation research, 196(2), 213-224.

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In vivo Xenograft Studies of IDH1-Mutant Cancers

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Female athymic nu/nu mice (Envigo/Harlan) were used for all in vivo xenograft studies. All studies were approved by the Yale University IACUC. Mice were quarantined for at least 1 week before experimental manipulation. Human luciferase-expressing HCT116 cells (with and without IDH1 mutation), HeLa cells (with and without IDH1 mutation), and HT1080 cells (with native IDH1 R132C mutation) were implanted subcutaneously (2 × 10⁶ cells in 0.1 cc PBS) in the right flank. Mice were visually observed daily, and tumors were measured three times per week by calipers to determine tumor volume using the formula:

V = \frac{4}{3} π \times (\frac{length}{2}) \times (\frac{width}{2}) \times (\frac{depth}{2})

. Olaparib (Selleckchem) was solubilized in DMSO and diluted with 10% (w/v) 2-hydroxy-propyl-beta-cyclodextrin (Sigma) to obtain the desired concentration. Olaparib was delivered via intraperitoneal injection (50 mg/kg) once daily, five days per week, starting four days after tumor implantation. Growth curves were compared statistically using ANOVA for repeated measures.

Sulkowski P.L., Corso C.D., Robinson N.D., Scanlon S.E., Purshouse K.R., Bai H., Liu Y., Sundaram R.K., Hegan D.C., Fons N.R., Breuer G.A., Song Y., Mishra-Gorur K., De Feyter H., de Graaf R.A., Surovtseva Y.V., Kachman M., Halene S., Günel M., Glazer P.M, & Bindra R.S. (2017). 2-Hydroxyglutarate produced by neomorphic IDH mutations suppresses homologous recombination and induces PARP inhibitor sensitivity. Science translational medicine, 9(375), eaal2463.

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4

Evaluating PEA15 Roles in Metastasis

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MDA-MB-231 PEA15-KO cells were used to establish stable cell lines overexpressing empty vector, PEA15-WT, PEA15-AA and PEA15-DD. Female athymic nu/nu mice, age 6 weeks, were purchased (Envigo). MDA-MB-231 PEA15-KO cells stably expressing PEA15 constructs (2 × 10⁶ cells/100 µL PBS) were injected into the tail veins of the mice. At 8 weeks after inoculation, animals were euthanized, and the lung tissues were analyzed for metastasis. The experimental metastasis model will allow us to determine the two late events of the metastatic process, extravasation and organ colonization.

Park J., Tacam M.J., Chauhan G., Cohen E.N., Gagliardi M., Iles L.R., Ueno N.T., Battula V.L., Sohn Y.K., Wang X., Kim H.S., Krishnamurthy S., Fowlkes N.W., Green M.M., Bartholomeusz G.A., Tripathy D., Reuben J.M, & Bartholomeusz C. (2021). Nonphosphorylatable PEA15 mutant inhibits epithelial-mesenchymal transition in triple-negative breast cancer partly through the regulation of IL-8 expression. Breast Cancer Research and Treatment, 189(2), 333-345.

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5

Athymic Nude Mouse Model Protocol

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Female athymic nu/nu mice were purchased from Envigo at age of 6–8 weeks. All animal experiments were conducted in accordance with the National Institutes for Health Guide for the Care and Use of Laboratory Animals and were approved by the University of Mississippi Medical Center’s Institutional Animal Care and Use Committee.

Dragojevic S., Ryu J.S., Hall M.E, & Raucher D. (2022). Targeted Drug Delivery Biopolymers Effectively Inhibit Breast Tumor Growth and Prevent Doxorubicin-Induced Cardiotoxicity. Molecules, 27(11), 3371.

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Xenograft Experiment with Breast Cancer in Mice

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Female nu/nu athymic mice were purchased from Envigo at the age of 4–6 weeks. Female mice were utilized since xenograft experiments were performed with breast cancer cell lines. The mice were randomly assigned to experimental groups and xenograft experiments were started 7–10 days after the mice were received. Mice were group housed (5 maximum) in conventional cages with unrestricted food and water access in a human xenograft designated area following animal biosafety level-2 procedures. The room was maintained at 21–23 °C and a 12 hours light-dark cycle. PicoLab Rodent Diet 5053 (Labdiet, Purina) containing 20% protein and 5% fat was used. Animal husbandry was carried out by Weill Cornell Medicine Belfer Research Building Vivarium technical staff. The mice were maintained and treated in compliance to Weill Cornell Medicine Institutional Animal Care and Use Committee protocols.

Gomes A.P., Ilter D., Low V., Rosenzweig A., Shen Z.J., Schild T., Rivas M.A., Er E.E., McNally D.R., Mutvei A.P., Han J., Ou Y.H., Cavaliere P., Mullarky E., Nagiec M., Shin S., Yoon S.O., Dephoure N., Massagué J., Melnick A.M., Cantley L.C., Tyler J.K, & Blenis J. (2019). Dynamic incorporation of histone H3 variants into chromatin is essential for acquisition of aggressive traits and metastatic colonization. Cancer cell, 36(4), 402-417.e13.

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7

Athymic Mice Xenograft Model

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Female nu/nu athymic mice (Envigo) were purchased at the age of 4–6 weeks, and the experiments were started 7–10 days after the mice were received at the Weill Cornell Medicine Belfer Research Building Vivarium. Experimental groups of 7–10 mice were created randomly and mice were group housed (maximum 5 in a cage) in standard cages with unrestricted acidified water and food, PicoLab Rodent Diet 5053 (Labdiet, Purina) containing 20% protein and 5% fat. Only deviation from the standard housing was for animals that received MMA in their drinking water as described in “Orthotopic Xenograft Experiments in Mice” section. Animal husbandry was carried out by the vivarium technical staff in a human xenograft designated area following animal biosafety level-2 procedures. The room was maintained at 21–23°C and a 12h light-dark cycle. The mice were maintained in compliance to Weill Cornell Medicine Institutional Animal Care and Use Committee protocols. The tumor size limit on the protocol was 20 mm on the largest dimension or 2.5 cm³ tumor volume or 10% of body weight, whichever was reached first. For mice studies no statistical method was used to predetermine sample size, mice were randomly distributed among the treatment groups and no blinding was performed.

Gomes A.P., Ilter D., Low V., Endress J.E., Fernández-García J., Rosenzweig A., Schild T., Broekaert D., Ahmed A., Planque M., Elia I., Han J., Kinzig C., Mullarky E., Mutvei A.P., Asara J., de Cabo R., Cantley L.C., Dephoure N., Fendt S.M, & Blenis J. (2020). Age-induced methylmalonic acid accumulation promotes tumor progression. Nature, 585(7824), 283-287.

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8

Xenograft Tumor Model in Nu/Nu Mice

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Female nu/nu athymic mice (Envigo) were purchased at the age of 6–7 weeks, and the experiments were started 7–10 days after the mice were received at the Weill Cornell Medicine Belfer Research Building Vivarium. Experimental groups of up to 10 mice were created randomly and mice were group housed (maximum five in a cage) in standard cages with unrestricted acidified water and food (PicoLab Rodent Diet 5053 (Labdiet, Purina) containing 20% protein and 5% fat). Animal husbandry was carried out by the vivarium technical staff in a human xenograft designated area following animal biosafety level-2 procedures. The room was maintained at 21–23 °C with a 12 h light–dark cycle. The mice were maintained in compliance with Weill Cornell Medicine Institutional Animal Care and Use Committee protocols. The tumor size limit on the protocol was 20 mm on the largest dimension or 2.5 cm³ tumor volume or 10% of body weight, whichever was reached first. For mouse studies, no statistical method was used to predetermine sample size, mice were randomly distributed among the treatment groups and no blinding was performed. All mouse studies have received ethical approval by IACUC at Weill Cornell Medicine under protocol number 2014-0060.

Li Z., Low V., Luga V., Sun J., Earlie E., Parang B., Shobana Ganesh K., Cho S., Endress J., Schild T., Hu M., Lyden D., Jin W., Guo C., Dephoure N., Cantley L.C., Laughney A.M, & Blenis J. (2022). Tumor-produced and aging-associated oncometabolite methylmalonic acid promotes cancer-associated fibroblast activation to drive metastatic progression. Nature Communications, 13, 6239.

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9

Athymic Mice Xenograft Model

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Female nu/nu athymic mice (Envigo) were purchased at the age of 4–6 weeks, and the experiments were started 7–10 days after the mice were received at the Weill Cornell Medicine Belfer Research Building Vivarium. Experimental groups of 7–10 mice were created randomly and mice were group housed (maximum 5 in a cage) in standard cages with unrestricted acidified water and food, PicoLab Rodent Diet 5053 (Labdiet, Purina) containing 20% protein and 5% fat. Only deviation from the standard housing was for animals that received MMA in their drinking water as described in “Orthotopic Xenograft Experiments in Mice” section. Animal husbandry was carried out by the vivarium technical staff in a human xenograft designated area following animal biosafety level-2 procedures. The room was maintained at 21–23°C and a 12h light-dark cycle. The mice were maintained in compliance to Weill Cornell Medicine Institutional Animal Care and Use Committee protocols. The tumor size limit on the protocol was 20 mm on the largest dimension or 2.5 cm³ tumor volume or 10% of body weight, whichever was reached first. For mice studies no statistical method was used to predetermine sample size, mice were randomly distributed among the treatment groups and no blinding was performed.

Gomes A.P., Ilter D., Low V., Endress J.E., Fernández-García J., Rosenzweig A., Schild T., Broekaert D., Ahmed A., Planque M., Elia I., Han J., Kinzig C., Mullarky E., Mutvei A.P., Asara J., de Cabo R., Cantley L.C., Dephoure N., Fendt S.M, & Blenis J. (2020). Age-induced methylmalonic acid accumulation promotes tumor progression. Nature, 585(7824), 283-287.

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Xenograft and Pancreatic Organoid Protocols

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Female nu/nu athymic mice were purchased from Envigo at the age of 4-6 weeks. Xenograft experiments were started 7-10 days after the mice were received. Kras +/LSL-G12D C57BL/6 mice were a kind gift from Dr. Lukas Dow, and were immediately used to isolate pancreatic organoids. The nu/nu mice were maintained at Weill Cornell Medicine in compliance with Weill Cornell Medicine Institutional Animal Care and Use Committee protocols.

Schild T., McReynolds M.R., Shea C., Low V., Schaffer B.E., Asara J.M., Piskounova E., Dephoure N., Rabinowitz J.D., Gomes A.P, & Blenis J. (2021). NADK is activated by oncogenic signaling to sustain pancreatic ductal adenocarcinoma. Cell reports, 35(11).

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Female nu nu athymic mice

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10 protocols using female nu nu athymic mice

Xenograft Study of ARRB2 Overexpression

Athymic Mouse Xenograft Model

In vivo Xenograft Studies of IDH1-Mutant Cancers

Evaluating PEA15 Roles in Metastasis

Athymic Nude Mouse Model Protocol

Xenograft Experiment with Breast Cancer in Mice

Athymic Mice Xenograft Model

Xenograft Tumor Model in Nu/Nu Mice

Athymic Mice Xenograft Model

Xenograft and Pancreatic Organoid Protocols

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