Animal procedures had approval of the Vanderbilt University Institutional Animal Care and Use Committee. For in vivo metastatic models, NOD-SCID-IL2Rγc (NSG) mice (6–8 weeks) (Jackson Laboratory) were injected in the left cardiac ventricle with 1×105 cells/100 μL PBS. Mice injected with MCF-7 cells received a 17β-estradiol pellet (0.36-mg 60-day release; Innovative Research of America). Mice bearing MCF-7 metastasis were injected IP with vehicle (10% (2-hydroxypropyl)-β-cyclodextrin (HPBCD) in 10% DMSO) or (1b ) (40 mg/kg) 2h prior to euthanasia (day 50). Mice injected with HDQ-P1-Luc were randomized and at 2 h after injection were treated for 5d with HPBCD, (1b ) (40 mg/kg) IP Q12h or trametinib (2 mg/kg, Santa Cruz Biotechnology, Inc) IP Q24 h. For bioluminescence imaging, mice were injected IP with RediJect D-Luciferin (1.5 mg) (PerkinElmer, Inc.) and imaged with a Xenogen IVIS using Living Image acquisition software (Xenogen Corp.). For ex vivo imaging, organs were placed in D-Luciferin (150 mg/mL PBS). After imaging tissue was fixed in 4% buffered formalin and paraffin-embedded.
Nod scid il2rγc nsg mice
Manufactured by Jackson ImmunoResearch
Sourced in United States
The NOD/SCID/IL2Rγc−/− (NSG) mouse is a highly immunodeficient mouse strain. It lacks mature T cells, B cells, and natural killer cells, making it a suitable model for the engraftment of human cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Taqman 48 system, by Roche (2 mentions)
Hiv 1 kit v 2, by Roche (2 mentions)
17β estradiol pellet, by Innovative Research (1 mentions)
Rediject d luciferin, by PerkinElmer (1 mentions)
Trametinib, by Santa Cruz Biotechnology (1 mentions)
Busulfan, by Merck Group (1 mentions)
Anti cd34 conjugated microbeads, by Miltenyi Biotec (1 mentions)
4 protocols using nod scid il2rγc nsg mice
1
Evaluation of Anti-Metastatic Drug Compounds
2
Humanized Mouse Model for HIV-1 Challenge
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Male 6–8-week-old NOD/SCID/IL2Rγc−/− (NSG) mice (Jackson Labs, Bar Harbor, ME, USA) were injected intramuscularly with NMCAB or CAB LAP at 45 mg CAB equivalents/kg. Eleven days after nanoformulation treatments, mice were reconstituted by intraperitoneal injection with 25 × 106 human peripheral blood lymphocytes (PBL) obtained by leukapheresis and centrifugal elutriation. Eleven days after reconstitution, mice were challenged with 104 50% tissue culture infectious dose (TCID50) HIV-1ADA by intraperitoneal injections. Mice were sacrificed 10 days after viral challenge. The experimental timeline is shown in Fig. 7A . Peripheral blood was collected at days 10 (prior to PBL reconstitution), 21 (prior to HIV-1 challenge), and 32 (10 days post HIV-1 challenge) for flow cytometry analysis of human pan-CD45, CD3, CD4 and CD8 immune markers [38 (link)]. Plasma was collected via centrifugation at 2000 × g for 5 min for drug quantitation by UPLC-MS/MS. HIV-1 RNA was analyzed in day 32 plasma samples using the Roche Amplicor and Taqman-48 system with HIV-1 kit V 2.0 according to the manufacturer’s instructions (Roche Diagnostics, Indianapolis, USA). Tissues were collected for CAB concentrations by UPLC/MS/MS, viral RNA and DNA quantitation by semi-nested real-time PCR [39 (link)], and immunohistochemical staining for HIV-1p24 antigen as described previously [40 (link)].
3
Humanized Mouse Model for HIV-1 Challenge
4
Engrafting Human HSPCs in NSG Mice
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NOD-scid-IL2Rγc−/− (NSG) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA) and housed in a specific pathogen-free facility. Eight-week-old female NSG mice were purchased and housed in a specific pathogen-free facility. For systemic CSE administration, 3R4F (0.5 mL of CSE/kg) was injected intravenously for 4 days, and the control group was injected with 100 µL PBS. To investigate the effect of CSE on the MSC niche, hUCB-derived CD34+ HSPCs were cocultured with CSE-treated hMSCs for 3 days and enriched by magnetic activated cell sorting using anti-CD34-conjugated microbeads (Miltenyi Biotec). For NSG bone marrow ablation, busulfan (Sigma‒Aldrich, St. Louis, MO, USA) was dissolved in dimethyl sulfoxide (DMSO, 16 mg/mL) and diluted with PBS at 1:4 ratio. Diluted busulfan was intraperitoneally injected into NSG mice (20 mg/kg). The next day, 5 × 104 CD34+ cells were suspended in 100 µL PBS and infused into the NSG mice via intravenous injection.
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