Nod cg prkdcscid j

Manufactured by Jackson ImmunoResearch

The NOD.Cg-Prkdcscid/J is a laboratory mouse strain that is deficient in functional T cells and B cells, resulting in a severe combined immunodeficiency (SCID) phenotype. This strain is commonly used in research as a model for studying cell transplantation, tumor growth, and other immunological processes.

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NOD.Cg-Prkdcscid/J (NOD/SCID, NOD.Cg-Prkdcscid/J) mice NOD/SCID mice (NOD.Cg-Prkdcscid/J

13 protocols using nod cg prkdcscid j

Targeting Leukemia Stem Cells in Murine CML and AML

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Twelve weeks-old NOD.Cg-Prkdc^scid/J (SCID) female mice were purchased from The Jackson Laboratory. SCID mice bearing GFP+ BCR-ABL1 CML-like or GFP+ FLT3(ITD);Tet2−/− AML-like murine leukemias (10%–20% GFP+ leukemia cells in total tail vein blood leukocytes) were treated for 7 consecutive days with vehicle, TKi (100 mg/kg imatinib or 1 mg/kg quizartinib) + PARPi (0.165 mg/kg talazoparib) as described before (Maifrede et al., 2018 (link); Nieborowska-Skorska et al., 2017b (link)), TGFβRi (10 mg/kg SB431542) (Shi et al., 2017 (link)) and the combination of TKi + PARPi + TGFβRi (Selleckchem). GFP+ leukemia cells were detected by flow cytometry in peripheral blood leukocytes (PBL), splenocytes (SPL) and bone marrow cells (BMC) 3 days after the end of treatment. Moreover, 1 × 10⁶ femoral bone marrow cells were transplanted into the sub-lethally irradiated secondary recipients to assess the effect of treatment on LSCs. Median survival time (MST) of the primary and secondary recipients was determined by Kaplan-Meier survival analysis. The Institutional Animal Care and Use Committee at Temple University approved all animal procedures and the experimental mice were maintained under a completely pathology-free condition and standard feeding diet in the animal facility of Temple University.

Le B.V., Podszywalow-Bartnicka P., Maifrede S., Sullivan-Reed K., Nieborowska-Skorska M., Golovine K., Yao J.C., Nejati R., Cai K.Q., Caruso L.B., Swatler J., Dabrowski M., Lian Z., Valent P., Paietta E.M., Levine R.L., Fernandez H.F., Tallman M.S., Litzow M.R., Huang J., Challen G.A., Link D., Tempera I., Wasik M.A., Piwocka K, & Skorski T. (2020). TGFβR-SMAD3 Signaling Induces Resistance to PARP Inhibitors in the Bone Marrow Microenvironment. Cell reports, 33(1), 108221.

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Mice Strains for Experimental Research

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C57BL/6J (JAX stock #000664), BALB/c (JAX stock #000651), NOD.Cg-Prkdc^scid/J (NOD-scid, JAX stock #001303), B6.Cg-Prkdc^scid/SzJ (B6-scid, JAX stock #001913) and NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ (NSG, JAX stock #005557) mice were obtained from the Jackson Laboratory. The mice were fed on a chow diet ad libitum and housed in a specific pathogen-free facility in plastic cages at 22 °C and 40–50% humidity, with a daylight cycle from 6 a.m. to 6 p.m. The animal protocols for the experiments described in this manuscript were approved by the Institutional Animal Care and Use Committee of The Jackson Laboratory. The mice were matched for age and sex in each experiment.

Li P., Lu M., Shi J., Hua L., Gong Z., Li Q., Shultz L.D, & Ren G. (2020). Dual roles of neutrophils in metastatic colonization are governed by the host NK cell status. Nature Communications, 11, 4387.

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Transgenic Mouse Strains for Immunological Research

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Mice were maintained on a 12 light/ dark cycle in a barrier facility, had ad libitum access to food and water, were exclusively used for the purposes of this study, and had not undergone any additional procedures. Mating cages housed one male and two females of the same genotype, and males were separated from dams at the time of delivery. Injections were performed on entire litters of 0–3-day-old mice of the following strains: C57BL/6J, Rag1^−/− (B6.129S7-Rag1^tm1Mom/J; strain 002216, Jackson laboratories), Cxcl10^−/− (B6.129S4-Cxcl10^tm1Adl/J; strain 006087, Jackson laboratories), CD4-deficient (B6.129S2-Cd4^tm1Mak/J; strain 002663, Jackson laboratories), CD8-deficient (B6.129S2-Cd8^atm1Mak/J; strain 002665, Jackson laboratories), CD4/CD8-deficient (intercrossed strains 002663 and 002665), NOD/SCID (NOD.Cg-Prkdc^scid/J; strain 001,303, Jackson laboratories), Ccl5^−/− (B6.129P2-Ccl5^tm1Hso/J; strain 005090; Jackson laboratories) and Cx3cr1^−/−; Ccr2^−/− [47 (link)] mice. Injected pups were allowed to recover and were subsequently immediately returned to their maternal cage until weaning. Mice of both sexes were randomly assigned to all experimental groups without bias, and the investigators were blinded until final data analysis during all of the experiments.

Anastasaki C., Chatterjee J., Cobb O., Sanapala S., Scheaffer S.M., De Andrade Costa A., Wilson A.F., Kernan C.M., Zafar A.H., Ge X., Garbow J.R., Rodriguez F.J, & Gutmann D.H. (2022). Human induced pluripotent stem cell engineering establishes a humanized mouse platform for pediatric low-grade glioma modeling. Acta Neuropathologica Communications, 10, 120.

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Athymic Nude and NOD.scid Mice Maintenance

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Athymic, nude (Foxn1^nu) or NOD.Cg-Prkdc^scid/J (NOD.scid) 6–8-week-old, female mice were obtained from the Jackson Laboratory and maintained in compliance with Institutional Animal Care and Use Committee (IACUC) guidelines. All animal care and protocols were in accordance with the Duke Animal Care and Use Program. Mice were group housed (3-5 mice per cage) in temperature controlled facility with a 12:12 h light:dark cycle. Food and water were provided ad libitum.

Bose S., Huang Q., Ma Y., Wang L., Rivera G.O., Ouyang Y., Whitaker R., Gibson R.A., Kontos C.D., Berchuck A., Previs R.A, & Shen X. (2022). G6PD inhibition sensitizes ovarian cancer cells to oxidative stress in the metastatic omental microenvironment. Cell reports, 39(13), 111012.

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Murine Lyme Disease Infection Model

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Five to eight-week-old female C3H/HeJ or NOD.Cg-Prkdc^scid/J (scid; Jackson Laboratories, Bar Harbor, ME) mice were needle-inoculated with 1 × 10⁴ organisms cultivated in vitro or from freshly harvested DMCs. At designated time points, animals were sacrificed, and blood and tissues collected for serology and culturing in BSK-II containing Borrelia antibiotic cocktail (0.05 mg/ml sulfamethoxazole, 0.02 mg/ml phosphomycin, 0.05 mg/ml rifampicin, 0.01 mg/ml trimethoprim and 2.5 μg/ml amphotericin B). Cultures were examined weekly by dark-field microscopy for up to 4 weeks. Seroconversion was determined by immunoblotting B. burgdorferi whole cell lysates with infected mouse serum, diluted 1:1000, followed by incubation with HRP-conjugated secondary antibody (Southern Biotechnology Associates, Birmingham, AL) diluted 1:20,000 and detection using SuperSignal West Pico chemiluminescence substrate (Pierce, Rockford, IL).

Groshong A.M., Grassmann A.A., Luthra A., McLain M.A., Provatas A.A., Radolf J.D, & Caimano M.J. (2021). PlzA is a bifunctional c-di-GMP biosensor that promotes tick and mammalian host-adaptation of Borrelia burgdorferi. PLoS Pathogens, 17(7), e1009725.

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Orthotopic Pancreatic Cancer Xenograft and 3-MA Treatment

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C57BL/6J and NOD.Cg-Prkdcscid/J (referred to as NOD mice) mice were obtained from The Jackson Laboratory. Young (8 weeks) and aged (78 weeks) male C57BL/6J mice (The Jackson Laboratory) were used for orthotopic implantations. In total, 0.5 × 10⁴ T4-KPC cells were injected into the mouse pancreas. After necropsy, tumor tissue and muscles were flash frozen in liquid nitrogen or formalin fixed for further analysis.
For 3-MA studies, 0.5 × 10⁴ T4-KPC cells were injected into the mouse pancreas. After 7 days of implantation, mice were randomized into 2 groups: vehicle or 3-MA–treated. 3-MA (30 mg/kg) was dissolved in saline and injected via i.p. to tumor-bearing mice once weekly. Saline was used as a vehicle control. Knockdown (RNA interference) studies: 0.5 × 10⁴ T4 shScr, T4 shPerp 14, and T4shPerp 146 were injected in 78-week-old C57BL/6J male mice. 3-MA treatment protocols were performed as described above.
Human pancreatic cancer organoids were a gift from Martin Fernandez-Zapico (Mayo Clinic). Approximately 100 organoids were injected in the pancreas of NOD mice. After 10 days of implantation, mice were randomized into 2 groups; the experimental group was i.p. injected weekly with 3-MA as described above, while the control group received saline (vehicle) injections.

Dasgupta A., Arneson-Wissink P.C., Schmitt R.E., Cho D.S., Ducharme A.M., Hogenson T.L., Krueger E.W., Bamlet W.R., Zhang L., Razidlo G.L., Fernandez-Zapico M.E, & Doles J.D. (2022). Anticachectic regulator analysis reveals Perp-dependent antitumorigenic properties of 3-methyladenine in pancreatic cancer. JCI Insight, 7(2), e153842.

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Athymic Nude and NOD.scid Mice Maintenance

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Preclinical Evaluation of Tumor Models

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All animal procedures were approved by the OMRF’s Institutional Animal Care and Use Committee. At the endpoint, animals were humanely euthanized by CO₂ inhalation as described in the approved IACUC animal use protocol (#22-01). Adult female, NOD/SCID mice (NOD.Cg-Prkdc^scid/J; The Jackson Laboratory; Strain #001303) were used for in vivo drug response studies and evaluation of tumor growth rate. Adult female, NRG mice (NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ; The Jackson Laboratory, Strain #007799) were used for PDX tumor growth rate and drug-response evaluation in vivo.

Drumond-Bock A.L., Wang L., Wang L., Cybula M., Rostworowska M., Kinter M, & Bieniasz M. (2023). Increased expression of BRD4 isoforms long (BRD4-L) and short (BRD4-S) promotes chemotherapy resistance in high-grade serous ovarian carcinoma. Genes & Cancer, 14, 56-76.

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Murine Strain Handling and Euthanasia

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Male and female (6–10 week) C57BL/6 J(Strain #:000664), NOD.Cg-Prkdc^scid/J(Strain #:001303), C57BL/6-Tg(TcraTcrb)1100Mjb/J (Strain #:003831), B6.129S7-Rag1^tm1Mom/J (Strain #:002216), B6.SJL-Ptprc^aPepc^b/BoyJ (Strain #:002014) mice were purchased from Jackson Laboratory. All mice were maintained in specific pathogen free animal facility with controlled temperature (68–79 ^oF), humidity (30–70%), and light/dark cycle (lights between 6 am and 6 pm). Mice were euthanized at the end of the experiment with CO₂ asphyxiation with cervical dislocation as a physical secondary assurance method. All animal experiments were performed with ethical compliance and approval from Institutional Animal Care and Use Committee of the University of Texas Southwestern Medical Center.

Feng Q., Liu Z., Yu X., Huang T., Chen J., Wang J., Wilhelm J., Li S., Song J., Li W., Sun Z., Sumer B.D., Li B., Fu Y.X, & Gao J. (2022). Lactate increases stemness of CD8 + T cells to augment anti-tumor immunity. Nature Communications, 13, 4981.

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NOD/SCID Mice Xenograft Protocol

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The immunodeficient NOD/SCID mice (NOD.Cg-Prkdc^Scid/J) were procured from The Jackson Laboratory and were maintained as per their recommended guidelines. All procedures in the mice xenograft studies were implemented in accordance with the guidelines of Institutional Animal Ethics Committee of the Indian Institute of Technology Kanpur, India and approved by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environment, Forest and Climate Change, Government of India.

Manzar N., Khan U.K., Goel A., Carskadon S., Gupta N., Palanisamy N, & Ateeq B. (2024). An integrative proteomics approach identifies tyrosine kinase KIT as a therapeutic target for SPINK1-positive prostate cancer. iScience, 27(3), 108794.

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