Five to six week old Nod-scid (NOD.CB17-Prkdcscid/J) and NOD-scid gamma (NOD-scid IL2Rgnull) mice were obtained from the Jackson Laboratories. 107 tumor cells were injected into the flanks in 0.9% sterile saline. Tumor and body weight was measured twice weekly. All studies beyond primary tumors (Figures 1A ) were on tumors reimplanted once and cell lines derived therefrom. All animal studies were SBP ACUC approved
Nod scid il2rgnull
Manufactured by Jackson ImmunoResearch
Sourced in United States
The NOD-scid IL2Rgnull is a laboratory mouse strain. It is deficient in the IL2 receptor gamma chain, leading to impaired immune function.
Automatically generated - may contain errors
Lab products found in correlation
Net en, by Merck Group (2 mentions)
C57bl 6j, by Jackson ImmunoResearch (2 mentions)
Nod scid gamma (nsg), by Jackson ImmunoResearch (2 mentions)
Nod cb17 prkdcscid j, by Jackson ImmunoResearch (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Ivis spectrum in vivo imaging system, by PerkinElmer (1 mentions)
Nod scid il2rgammanull, by Jackson ImmunoResearch (1 mentions)
Cd45.1 c57bl 6, by Jackson ImmunoResearch (1 mentions)
12 protocols using nod scid il2rgnull
1
Xenograft Tumor Implantation in NOD-scid Mice
2
Standardized Mouse Model for Contraceptive Treatments
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All procedures were approved by the Ohio State University Institutional Animal Care and Use Committee and the Stanford University Administrative Panel on Laboratory Animal Care. As indicated, 6– to 8–wk-old C57BL/6J (wild-type) and NOD-scid IL-2Rgnull (NSG) female mice were purchased from Jackson Laboratory (Bar Harbor, ME). As indicated, mice were s.c. injected for five consecutive days with 100 μl of PBS alone or a PBS solution containing 1 mg of NET-EN (Sigma-Aldrich, Laramie, WY) (prepared by glass-to-glass homogenization and sonication), whereas other mice were s.c. administered a single dose of 1 mg of DMPA (Depo-Provera) or 2 mg of methylprednisolone (MePRDL) (Depo-Medrol) (both Pfizer, New York, NY). Where indicated, cells collected by vaginal lavage were stained with crystal violet to identify wild-type and NSG mice in the estrus stage of the estrous cycle (26 ). Also as indicated, mice were euthanized 24 h after the fifth NET-EN treatment, and the serum was sent to the Endocrine Technologies Support Core at the Oregon National Primate Research Center to quantify norethisterone (NET) levels by liquid chromatography–tandem triple quadrupole mass spectrometry (27 (link)).
3
Teratoma Formation from iPSCs
4
Animal Research on C57BL/6 and NSG Mice
5
Bioluminescent Tumor Monitoring in NSG Mice
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105-106 modified HeyA8 (always infected with a luciferase lentivirus) were injected i.p. into 6-week-old female NSG mice (NOD scid gamma, NOD-scid IL2Rgnull, NOD-scid IL2Rgammanul) (Jackson Laboratory, Cat.No.: 005557) following the Northwestern University Institutional Animal Care and Use Committee (IACUC)-approved protocol. The growth of tumor cells in the mice over time was monitored non-invasively using the IVIS® Spectrum in vivo imaging system (Perkin Elmer). The tumor load was quantified by the luminescence of the regions of interest (the same area for each mouse encompassing the entire abdomen) using the Living Image software. To visualize the tumor load in the mice, 200 µl of the sterile luciferin stock solution (15 mg/ml in Dulbeccos’s phosphate buffered saline without Mg2+ and Ca2+) was i.p. injected per mouse (which equals approximately 10 µl of luciferin stock per g of body weight). The mice were imaged 15 minutes after injection. Total Flux values of the different treatment groups were compared.
6
Rapid Orthotopic Implantation of Patient-Derived Tumors
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The patient tumor sample received through consent was immediately dissociated and prepared for injection. To reduce the lag time between the surgical removal of the patient tumor sample and the orthotopic injection into the mouse brain, cells were not counted. The prepared tumor sample was directly implanted into the brains of NSGTM mice (NOD-scid IL2Rgammanull, NOD-scid IL2Rgnull, NOD scid gamma, Jackson Laboratory, Bar Harbor, ME, USA). The injection site was 2 mm right, 1 mm anterior to the bregma suture. One µL of the prepared tumor sample was injected each minute for a total of 3 min. Mice were monitored daily for the first week after surgery for recovery. Mice were then assessed 3 times per week for signs of illness associated with tumor growth. Brains were collected from mice euthanized at morbidity. Frozen tissue was stored at −80 °C, for protein or nucleic acid preparations. PFA-preserved tissue was processed and embedded in paraffin. Five micrometer sections were cut for histology and IF-FISH.
7
Standardized Mouse Model for Contraceptive Treatments
8
Xenogeneic Graft-versus-Host Disease Induction
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Xenogeneic graft-vs-host-disease (xGVHD) was induced in female NSG mice (NOD-scidIL2RGnull, strain no. 005557, Jackson Laboratory), as described previously (39 (link), 40 (link)). Briefly, ex vivo-purified human naïve CD8+ T-cells were first cultured for 7 days in Tc0 and Tc1 differentiation cultures, as described above. On day 7, Tc cells were washed, resuspended with PBS, and injected intravenously into 2 Gy irradiated 6–8-week- old female NSG mice admixed with ex vivo-sorted allogeneic bulk CD4+ CD25- T-cells at a 1:1 ratio (10 × 106 CD4 and 10 × 106 CD8 cells/mouse) in the indicated groups. 10 × 106 CD4+ CD25- T-cells/mouse without CD8+ T-cells (1:0 ratio) served as the control group. Some mice also received ex vivo-purified bulk CD8+ T-cells (Miltenyi Biotech, 130-045-201) or in vitro-differentiated Tc0 or Tc1 cells alone (10 × 106 cells/mouse, 0:1 ratio), representing CD8+ T-cell only groups. Mice were monitored for weight loss up to 15 days post injection. Number of mice per experimental group per experimental replicate is indicated in the figure legend.
9
Isolation and Transplantation of Human Islets
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Human islets were obtained from the Integrated Islet Distribution Program. Human islets were processed for DA-ZP1 treatment or transplantation. Joslin Diabetes Center Institutional Review Board declared studies on de-identified autopsy tissue does not qualify as human subject research. Upon receipt, islets were centrifuged at 200g for 1 min and resuspended in Miami media (Mediatech). Islets were then transferred to 10 cm culture dishes and cultured overnight to 24 h. Healthy islets were handpicked and washed with DPBS. For transplantation experiments 500 IEQ were transplanted under the kidney capsule of NSG (NOD-scid-IL2Rgnull; Jackson Laboratory) mice as previously described (Kahraman et al, 2011 (link)). For DA-ZP1 staining, islets were incubated in culture medium containing DA-ZP1 for 30 min at 37°C. At the end of incubation time, cells were washed and cell pellet was resuspended in DA-ZP1 free Miami media. Donor demographic information is summarized in Table S2.
Table S2
10
Xenotransplantation of Purified EPCR+ Cells
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All experiments with animals were conducted under protocols approved by the University of Montreal Animal Care Committee. EPCR cell subsets purified from uncultured or expanded CD34+CD45RA- CB cells were transplanted by tail vein injection into sub-lethally irradiated (250 cGy, <24 hr before transplantation) 8 to 16-week-old female NSG (NOD-Scid IL2Rgnull, Jackson Laboratory) mice. Human cells NSG-BM cells were collected by femoral aspiration or by flushing the two femurs, tibias and hips when animals were sacrificed at week 26.
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