Nod cg prkdcscid il2rgtm1wjl szj

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The NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ is a mouse strain. It is a severe combined immunodeficiency (SCID) model with a targeted mutation in the Il2rg gene, which encodes the common gamma chain of the interleukin-2 receptor.

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310 protocols using nod cg prkdcscid il2rgtm1wjl szj

Intracranial GSC Xenografts in NSG Mice

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Intracranial xenografts were created by implanting 10,000
human-derived GSCs into the right cerebral cortex of NSG (NOD.Cg-Prkdcscid
Il2rgtm1Wjl/SzJ, The Jackson Laboratory, Bar Harbor, ME, USA) mice at a
depth of 3.5 mm. All mouse experiments were performed under an animal
protocol approved by the University of California, San Diego Institutional
Animal Care and Use Committee. Healthy, wild-type female mice of NSG
(NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ, The Jackson Laboratory, Bar Harbor, ME,
USA) background, 4–6 weeks old, were randomly selected and used in
this study for intracranial injection. Mice had not undergone prior
treatment or procedures. Mice were maintained in 14 hours light/10 hours
dark cycle by animal husbandry staff at the University of California, San
Diego, with no more than 5 mice per cage. Housing conditions and animal
status were supervised by a veterinarian. Animals were monitored until
neurological signs were observed, at which point they were sacrificed.
Neurological signs or signs of morbidity included hunched posture, gait
changes, lethargy and weigh loss. In parallel survival experiments, mice
were observed until the development of neurological signs. Healthy female
mice of NSG, 4–6 weeks old, were randomly selected and used in this
study for intracranial injection.

Xie Q., Wu T.P., Gimple R.C., Li Z., Prager B.C., Wu Q., Yu Y., Wang P., Wang Y., Gorkin D.U., Zhang C., Dowiak A.V., Lin K., Zeng C., Sui Y., Kim L.J., Miller T.E., Jiang L., Lee-Poturalski C., Huang Z., Fang X., Zhai K., Mack S.C., Sander M., Bao S., Kerstetter-Fogle A.E., Sloan A.E., Xiao A.Z, & Rich J.N. (2018). N6-methyladenine DNA Modification in Glioblastoma. Cell, 175(5), 1228-1243.e20.

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Xenograft Mouse Models for Cancer Research

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Animals were housed in cages of up to 5 mice in pathogen-free conditions at a barrier facility at the Lokey Stem Cell Building (SIM1) at Stanford School of Medicine (Stanford, California) or at the Children’s Hospital of Philadelphia Animal Facility, fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). All animal handling, surveillance, and experimentation was performed in accordance with and approval from the Stanford University Administrative Panel on Laboratory Animal Care (Protocol #APLAC-31287) or approved by the CHOP Institutional Animal Care and Use Committee (IACUC; Approved IACUC Protocol #643). For para-orthotopic and metastatic xenograft models, 6–10 weeks old male or female in-house bred NSG mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ: The Jackson Laboratory) were used. Animals bearing engrafted tumors were randomized into cohorts to ensure a similar mean tumor burden/group based on bioluminescent FLUX [P/s] values at study enrollment.
For patient derived xenograft models, 6–7 weeks old female (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ, purchased from: The Jackson Laboratory, #005557) mice were used. Animals bearing engrafted tumors were randomized assigned into cohorts, to ensure a similar mean tumor volume/group based on caliper measurements at study enrollment.

Heitzeneder S., Bosse K.R., Zhu Z., Zhelev D., Majzner R.G., Radosevich M.T., Dhingra S., Sotillo E., Buongervino S., Pascual-Pasto G., Garrigan E., Xu P., Huang J., Salzer B., Delaidelli A., Raman S., Cui H., Martinez B., Bornheimer S.J., Sahaf B., Alag A., Fetahu I.S., Hasselblatt M., Parker K.R., Anbunathan H., Hwang J., Huang M., Sakamoto K., Lacayo N.J., Klysz D.D., Theruvath J., Vilches-Moure J.G., Satpathy A.T., Chang H.Y., Lehner M., Taschner-Mandl S., Julien J.P., Sorensen P.H., Dimitrov D.S., Maris J.M, & Mackall C.L. (2021). GPC2-CAR T Cells Tuned for Low Antigen Density Mediate Potent Activity Against Neuroblastoma Without Toxicity.. Cancer cell, 40(1), 53-69.e9.

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Subcutaneous Tumor Implantation in NSG Mice

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All experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Georgia Regents University. Six-week-old female NSG (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ) mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained in accordance with IACUC guidelines. The animal experiments were performed using the NSG mouse model as described previously ^{22 (link)}. Briefly, 2×10⁶ cells, in 100 μl of Matrigel (BD Biosciences, Franklin Lakes, NJ, USA), were injected subcutaneously into the fourth abdominal fat pad at the base of the nipple. Tumor growth was measured externally every 3 to 5 days using vernier calipers and the tumor volume was calculated as length×width²×0.52. The mice were sacrificed on day 60, and the lungs were removed and processed for histological analyses. H&E staining was used to examine tumor metastasis in the lung.

Teng Y., Ren X., Li H., Shull A., Kim J, & Cowell J.K. (2015). Mitochondrial ATAD3A combines with GRP78 to regulate the WASF3 metastasis-promoting protein. Oncogene, 35(3), 333-343.

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Spontaneous Progression of Pancreatic Cancer in Mice

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Congenitally athymic nude mice (NCr-nu/nu) and mice NOD scid gamma (NSG^TM; NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ, (6–8-weeks old) were bred in-house or purchased from Jackson Laboratories (Strain# 005557). C57BL/6J mice (6–8-weeks old) were bred in-house. Mice were housed under pathogen-free conditions in a 12-h dark/12-h night (6:00 to 18:00) cycle at 70 °F with a humidity of ~50%. All animal experiments were performed with the approval of The University of Nebraska Medical Center Institutional Animal Care and Use Committee (IACUC).
Tumour tissues from 20–22-week-old female mice from the Kras^LSL.G12D/+; p53^R172H/+; Pdx1-Cre^tg/+ (KPC) mouse spontaneous progression model of pancreatic cancer on a C57BL/6 background and littermate controls were collected by euthanizing the mice at the respective ages.

Murthy D., Attri K.S., Shukla S.K., Thakur R., Chaika N.V., He C., Wang D., Jha K., Dasgupta A., King R.J., Mulder S.E., Souchek J., Gebregiworgis T., Rai V., Patel R., Hu T., Rana S., Kollala S.S., Pacheco C., Grandgenett P.M., Yu F., Kumar V., Lazenby A.J., Black A.R., Ulhannan S., Jain A., Edil B.H., Klinkebiel D.L., Powers R., Natarajan A., Hollingsworth M.A., Mehla K., Ly Q., Chaudhary S., Hwang R.F., Wellen K.E, & Singh P.K. (2024). Cancer-associated fibroblast-derived acetate promotes pancreatic cancer development by altering polyamine metabolism via the ACSS2–SP1–SAT1 axis. Nature Cell Biology, 26(4), 613-627.

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Subcutaneous Tumor Grafting in NSG Mice

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NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) mice were purchased from Jackson laboratories. Mice were group-housed (up to 5 per cage) in individually ventilated cages with ad libitum access to food and acidified water (pH 2.5 to 2.8) in a temperature (22.2 ± 0.5 °C) and humidity (30–70%) controlled facility with 12/12-h light/dark cycle. The animal care and use program is accredited by AAALAC. All animal experiments were approved by the Weill Cornell Institutional Animal Care and Use Committee. For all mouse studies, mice of either sex were used, and mice were randomly allocated to experimental groups, but blinding was not performed. Mice (aged 5–8 weeks) were age-matched for tumor inoculation. Group sizes were selected on the basis of prior knowledge. For subcutaneous grafting, sgCtrl, sgK#1 or #2-transduced Saos2 cells were resuspended in 50% Matrigel (BD Bioscience, #356231) in PBS and ~5,000,000 cells were injected into each flank of NSG mice. Tumor growth was monitored and measured every 7 days by caliper, and volume was calculated by the formula:

V = \frac{4 π}{3} * \frac{a}{2} * \frac{b}{2} * \frac{c}{2}

(V tumor volume; a tumor length; b tumor width; c tumor height). The endpoints were determined on the basis of the level of animal discomfort and tumor sizes. The maximum allowable tumor size is 20 mm in diameter.

Li F., Wang Y., Hwang I., Jang J.Y., Xu L., Deng Z., Yu E.Y., Cai Y., Wu C., Han Z., Huang Y.H., Huang X., Zhang L., Yao J., Lue N.F., Lieberman P.M., Ying H., Paik J, & Zheng H. (2023). Histone demethylase KDM2A is a selective vulnerability of cancers relying on alternative telomere maintenance. Nature Communications, 14, 1756.

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PDX Xenograft Tumor Imaging and Therapy

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The Memorial Sloan Kettering Institutional Animal Care and Use Committee approved all animal protocols for this work. Female NSG mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ; The Jackson Laboratory) that were 6 to 8 weeks old at time of patient-derived xenograft (PDX) implantation were used for all PET imaging and in vivo talazoparib efficacy experiments. The PDX SCRX-Lu149 was isolated and passaged as previously described^{21 (link),22 (link)}. PDXs were injected in the right shoulder for PET/CT experiments and in the right flank during tumor growth inhibition experiments.

Laird J., Lok B.H., Carney B., Kossatz S., de Stanchina E., Reiner T., Poirier J.T, & Rudin C.M. (2019). PET Imaging of a [18F]-radiolabeled PARP Inhibitor Monitors the Therapeutic Efficacy of Talazoparib in Small Cell Lung Cancer Patient-Derived Xenografts. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 14(10), 1743-1752.

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Assessing Engineered T Cell Efficacy in MDA-MB-468 Xenograft Mice

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Six to eight-week-old female NSG mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA) were injected with 5 × 10⁶ IL4 cytokine-producing, MUC1-overexpressing MDA MB 468 (MDA MB 468/IL4) cells suspended in 50%DPBS/50%matrigel subcutaneously (s.c.) into the left inferior mammary fat pad. Once the tumor reached a size of approx. 75 mm³ (~ 4–5 weeks), animals were injected intravenously (i.v.) with 3 × 10⁶ eGFP-FFLuc+1G, 1G.4/7ICR, 2G, or 2G.4/7ICR T cells. Tumor size was measured by bi-weekly caliper measurement and tumor volume (mm³) was calculated by length x width x width/2. T cell expansion and persistence was monitored using the IVIS Lumina In Vivo Imaging system (Caliper Life Sciences, Hopkinton, MA) 10 min after injection (i.p.) with 100ul of D-luciferin (15 mg/mL). All in vivo analysis was performed using Living Image software (Caliper Life Sciences, Inc., Hopkinton, MA). Experiments were performed according to Baylor College of Animal Husbandry guidelines.

Bajgain P., Tawinwung S., D’Elia L., Sukumaran S., Watanabe N., Hoyos V., Lulla P., Brenner M.K., Leen A.M, & Vera J.F. (2018). CAR T cell therapy for breast cancer: harnessing the tumor milieu to drive T cell activation. Journal for Immunotherapy of Cancer, 6, 34.

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Murine Xenograft Tumor Modeling

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For in vivo experiments, we used 6- to 12-week-old female nonobese diabetic severe combined immunodeficiency, interleukin 2 receptor gamma chain deficient, NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ (NSG) mice or BALB/cJ mice (The Jackson Laboratory, Bar Harbor, ME, USA). Animals were maintained in a specific pathogen-free facility at the University of British Columbia (UBC) Biomedical Research Centre. All experiments were conducted with approval of the UBC Animal Care Committee.
Primary tumor development was examined following subcutaneous (s.c.) injection of MDA-MB-231 cells (1 × 10⁶) prepared in BD Matrigel™ (BD Biosciences, San Jose, CA, USA) into the right hind flank of NSG mice. Tumor growth was measured using manual calipers, and the tumor volume was estimated using the following formula: length times width² divided by 2. Final tumor masses were measured after excision and the tumors were retained for histochemical analyses. Flow cytometry was performed on lung digests to enumerate tumor cells based on detection of GFP or RFP fluorescence.

Snyder K.A., Hughes M.R., Hedberg B., Brandon J., Hernaez D.C., Bergqvist P., Cruz F., Po K., Graves M.L., Turvey M.E., Nielsen J.S., Wilkins J.A., McColl S.R., Babcook J.S., Roskelley C.D, & McNagny K.M. (2015). Podocalyxin enhances breast tumor growth and metastasis and is a target for monoclonal antibody therapy. Breast Cancer Research : BCR, 17(1), 46.

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Lentiviral CDC20 Knockdown Glioma Model

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TICs were transduced with lentiviral vectors expressing CDC20 or a non-targeting control (shCONT) shNRA for the knockdown experiments. 36 hours post infection, viable cells were counted and engrafted intracranially into NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ, The Jackson Laboratory, Bar Harbor, ME) mice under a Cleveland Clinic Foundation Institutional Animal Care and Use Committee approved protocol. Animals were then maintained until neurological signs were apparent, at which point they were sacrificed. The brains were harvested and fixed in 4% formaldehyde, cryopreserved in 30% sucrose, and then cryosectioned. Sections were stained with hematoxylin and eosin. In parallel survival experiments, animals were monitored until they developed neurological signs.

Xie Q., Wu Q., Mack S.C., Yang K., Kim L., Hubert C.G., Flavahan W.A., Chu C., Bao S, & Rich J.N. (2015). CDC20 maintains tumor initiating cells. Oncotarget, 6(15), 13241-13254.

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Biodistribution of 177Lu-NNV003 in Xenograft Models

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Biodistribution of ¹⁷⁷Lu-NNV003 was performed in female NSG mice (NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ; The Jackson Laboratory, USA, 6–7 weeks old, average weight 20.6 g) with MEC-2 subcutaneous (s.c.) xenografts, and in female and male CB17 SCID mice (CB-17/Icr-Prkdc^scid/scid/Rj, Janvier Labs, France, 10 weeks old, average weight 21.7 g) with DOHH-2 s.c. xenografts. Mice were injected with 10 × 10⁶ MEC-2 cells or 5 × 10⁶ DOHH-2 cells in each flank. Mice were injected with 1 MBq (MEC-2) or 1.2 MBq (DOHH-2) of ¹⁷⁷Lu-NNV003 in 100 µl formulation buffer when tumours were of suitable size. The day prior to injection of RIC, 200 μg mouse IgG2a-κ (M7769-5MG, Sigma-Aldrich, USA) was administered intraperitoneally (i.p.). Mice were necropsied at different time points after RIC injection. Radioactivity was measured by gamma counters (Wizard 3470, PerkinElmer, USA or Cobra II auto-gamma detector, Packard Instrument Company, USA). Samples from the injected solution of ¹⁷⁷Lu-NNV003 were used as references in the measurement procedures.

Maaland A.F., Heyerdahl H., O’Shea A., Eiriksdottir B., Pascal V., Andersen J.T., Kolstad A, & Dahle J. (2019). Targeting B-cell malignancies with the beta-emitting anti-CD37 radioimmunoconjugate 177Lu-NNV003. European Journal of Nuclear Medicine and Molecular Imaging, 46(11), 2311-2321.

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