All mice were bred and housed in specific pathogen-free conditions in the animal barrier facility at the Cincinnati Children’s Hospital Medical Center (CCHMC). All animal studies were conducted in accordance with an approved Institutional Animal Care and Use Committee protocol and federal regulations. Lgr6−/− mice (Jackson stock #016934), NRG mice (NOD-Rag1null IL2rgnull, NOD rag gamma), Rag1−/− mice (Jackson stock #002216), C57BL/6 mice (Jackson stock #000664), C57BL/6 congenic BoyJ mice were purchased from Jackson or Comprehensive Mouse and Cancer Core of CCHMC. All BoyJ mice used are confirmed with the expression of NKp46 by flow cytometry analysis with peripheral blood samples. The MycG/G mice were a kind gift from Dr. H. Leighton Grimes at Cincinnati Children’s Hospital Medical Center, OH, USA. Mycf/f mice and Ncr1Cre mice were backcrossed to C57BL/6 background at our lab. All mice used were 8 to 12 weeks old. Age and sex matching were performed for each independent experiment. The MycG/G mice, MycΔ/Δ/Ncr1Cre mice were born at the expected Mendelian ratios and showed normal WBC, hemoglobin, and platelet counts.
Nrg mice
Manufactured by Jackson ImmunoResearch
NRG mice are genetically engineered mouse models that express the NRG gene. NRG mice are used for research purposes in the field of immunology and neurology.
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Lab products found in correlation
Rag1 mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Jackson ImmunoResearch (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Fulvestrant, by Selleck Chemicals (1 mentions)
Birinapant, by MedChemExpress (1 mentions)
Shiverer mice, by Jackson ImmunoResearch (1 mentions)
Gammacell 3000 elan, by Best Theratronics (1 mentions)
8 protocols using nrg mice
1
Genetically Modified Mice for Immunology
2
PDX Tumor Regression with Targeted Therapies
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Female 5–6-week-old NRG mice (Jackson labs stock #007799) were implanted with HCI-011 patient-derived xenograft (PDX) tumor fragments using a previously described protocol with estrogen supplementation (42 (link)). When tumors reached approximately 100 mm3, mice were randomized into treatment or control groups. Mice received either vehicle control (15% captisol IP, 3×/week for 5 weeks), birinapant (20 mg/kg IP, 3×/week for 5 weeks), fulvestrant (200 mg/kg subQ, 1×/week for 5 weeks), or the combination of birinapant and fulvestrant at the same doses as single agents. For birinapant treatment, birinapant (Medchem Express, HY-16591) was dissolved in 15% captisol at a concentration of 2 mg/mL (made fresh every time), and 100 μL/10 g mouse body weight was injected intraperitoneally. For fulvestrant treatment, 250 mg of fulvestrant (Selleck Chemicals, S1191) was dissolved in 0.5 mL DMSO, incubated at 37°C for 15 minutes and then diluted 1:20 with corn oil to give a final concentration of fulvestrant of 25 mg/mL (made fresh every time). 80 μL/10 g mouse body weight was injected subcutaneously. Animal experiments were all conducted in compliance with institutional guidelines and regulations after approval from the University of Utah Institutional Animal Care and Use Committee, under protocol 21-06008.
3
Analyzing GrA Production in GVHD
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To examine GrA production from human CD4+ T cells in the context of GVHD, NRG mice (originally from The Jackson Laboratory, purchased from the Purdue Biological Evaluation Core) were injected i.v. with 5 × 106 to 20 × 106 human PBMCs that had been recovered the previous day (37°C, 5% CO2, complete RPMI) from frozen stocks isolated from 2 distinct donors (from ZenBio). After injection, mice were monitored daily until they reached 20% body weight loss or developed a significant clinical score (>3). Upon euthanization, immune cells were isolated from the spleen, liver, SI, and LI; were stained for cell surface expression of human CD3, CD4, and CD8; and were fixed, permeabilized, and stained for intracellular human GrA, GrB, and perforin (see below). The frequency of GrA-, GrB-, and perforin-expressing T cells was subsequently determined by flow cytometry.
4
Preclinical Myelination Disorder Models
5
Humanized Mouse Model for HCMV Study
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NRG mice were obtained from The Jackson Laboratory (JAX, Bar Harbor, ME) and bred in-house under pathogen-free conditions. Female mice were used for all experiments (19). Briefly 5 weeks-old mice were sub-lethally irradiated (450 cGy) using a [137Cs] column irradiator (Gammacell 3000 Elan; Best Theratronics, Ottawa, Canada). Four hours after irradiation, 2.0x105 human CB-CD34+ cells were injected as described before for HSCT [19 (link)]. CB units were tested prior to experiments for humanization potential. Only CB units which resulted into >20% hCD45 (BL) at week 10 post-HCT were used in studies. Cryopreserved and quality-controlled iDCgB were thawed and injected into NRG at weeks 6, 7, 10 and 11 after HCT. Mice were immunized with a total dose of 5.0x105 iDCgB cells injected s.c., near the anatomical regions of the inguinal and axillary lymph nodes. Humanized NRG mice were infected with HCMV as described before (16). Briefly, at week 17 post HCT, mice were treated s.c. with 150 ng hG-CSF/mouse/day (Granocyte, Kohlpharma GmbH, Merzig, Germany) for 3 days, injected i.p. with 1.0x106 MRC-5 cells previously infected with TB40-GLuc (MOI 1) and treated for G-CSF for additional 2 days. For HCMV reactivation, mice were treated s.c. with 2,5 μg hG-CSF/mouse/day for 7 days from weeks 24 and 25 post-HCT.
6
Xenograft Tumor Model in Immunocompromised Mice
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Severely immunocompromised NRG mice (The Jackson Laboratory stock #007799) 10-16 weeks old (male or female) were used for all tumor implantations. Tumor volume was measured with digital calipers by taking 3 perpendicular measurements of LxWxH. Mice were monitored 2x/week for tumor growth and weight gain/loss and were euthanized when tumor volume reached 2000 mm3 or if other humane criteria were met. H69AR cells still in log-phase growth were harvested from culture flasks and injected subcutaneously in the flank at 5x106 cells/mouse using a 25 ga needle. Cells were mixed 1:1 with Matrigel (50 μL:50 μL) for injection in groups of 5-6 mice. PDX tumors were implanted subcutaneously in the flank after harvesting tumors from donor mice. Donor tumors were minced into a paste with a razor blade, large fibrous chunks were removed, and remaining paste was mixed 1:1 with Matrigel (50 μL:50 μL) for injection with 14 ga needles in groups of 5-6 mice.
7
Establishing NRG Mouse Colony
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NRG mice were purchased from the Jackson Lab (Bar Harbor, ME). The mice were bred and housed under specific pathogen-free conditions in a level II facility at the Central Animal Facility at McMaster University. Mouse colonies were maintained on a 12 h light/12 h dark cycle.
8
Humanized Mouse Model Sources
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