NOD/Shi-scid/IL-2Rγnull (NOG) mice were purchased from Taconic (Germantown, NY, USA) and kept in pathogen-free conditions in the animal facility at the Interdepartmental Centre of Experimental Research Service (CIRSAL) of the University of Verona, as approved by the Italian Health Ministry (Autorizzazione n°1294/2015-PR). U937 AML cell line (1 × 106) was injected into the tail vein of totally irradiated (1.2 Gy, 137Cesium source), 8–12-week-old mice. At day 9 post-injection, mice were assigned to one of the following treatment arms: DMSO or Ara-C (100 mg/kg), all administered through daily intraperitoneal injection for 3 days (from day +9 to +11). On day +12, each treatment arm was split into three arms: GSI-IX (10 mg/kg) or GSI-XII (10 mg/kg) or their vehicle (DMSO). Animals were sacrificed after 3 weeks from cell line injection, and bone marrow leukemic burden was evaluated as the percentage of human CD45+ cells. For survival assays, animals were sacrificed when body weight loss was equal to 20% according to ethical regulation.
Nod shi scid il 2rγnull nog mice
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The NOD/Shi-scid/IL-2Rγnull (NOG) mice are a strain of immunodeficient mice. They have a severe combined immunodeficiency (SCID) mutation and lack the interleukin-2 receptor gamma chain, which results in the absence of functional T cells, B cells, and natural killer cells.
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5 protocols using nod shi scid il 2rγnull nog mice
1
Xenograft Model of Acute Myeloid Leukemia
2
Orthotopic Liver Tumor Xenograft Model
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In vivo studies were performed in female NOD/Shi-scid/IL-2Rγnull (NOG) mice (Taconic Biosciences, Hudson, NY). 2 × 106 HepG2 and Huh-6 cells transduced with luciferase and resuspended in 100 μl phosphate-buffered saline (PBS) were surgically implanted into either the right lobe of the liver through a right flank incision or the left lobe with a midline, abdominal incision. The mice underwent BLI beginning at 10 days after implantation and every week thereafter with the In Vivo Imaging System (IVIS, PerkinElmer, Waltham, MA), and luminescence flux was recorded to assess tumor growth. After seven weeks (HepG2) or four weeks (Huh-6), necropsy was performed, intrahepatic and extrahepatic sites of tumor were noted, and samples were harvested for immunohistochemistry and RNA isolation.
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In vivo studies were performed in female NOD/Shi-scid/IL-2Rγnull (NOG) mice (Taconic Biosciences, Hudson, NY). 2 × 106 HepG2 and Huh-6 cells transduced with luciferase and resuspended in 100 μl phosphate-buffered saline (PBS) were surgically implanted into either the right lobe of the liver through a right flank incision or the left lobe with a midline, abdominal incision. The mice underwent BLI beginning at 10 days after implantation and every week thereafter with the In Vivo Imaging System (IVIS, PerkinElmer, Waltham, MA), and luminescence flux was recorded to assess tumor growth. After seven weeks (HepG2) or four weeks (Huh-6), necropsy was performed, intrahepatic and extrahepatic sites of tumor were noted, and samples were harvested for immunohistochemistry and RNA isolation.
3
NOG Mouse Hepatocellular Carcinoma Model
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All animal procedures used in this study were performed under an animal protocol approved by the Institutional Care and Use Committee of Baylor College of Medicine (AN-6191). In vivo studies were performed in male and female NOD/Shi-scid/IL-2Rγnull (NOG) mice (Taconic Biosciences, Hudson, NY, USA). 1.5×106 HepT1 cells transduced with lentiviral psi-LVRU6MP-GFP-mCherry and resuspended in 100 μl phosphate-buffered saline (PBS) were injected into the tail vein. After 12 to 13 weeks, necropsy was performed, and liver and lung samples were imaged and harvested for immunohistochemistry.
4
Macitentan Inhibits Myeloma Growth
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NOD/Shi-scid/IL-2Rγnull (NOG) mice were purchased from Taconic (Germantown, NY, USA). Four groups of five four-to six-week-old female mice were subcutaneously injected with 5 × 106 RPMI 8226 cells. At day +7 post-injection, mice were daily treated with macitentan (30 mg/kg) (21 (link)) or vehicle (Methocell 0.05% and TWEEN 80 0,05%) per os for four weeks. At the end of treatment mice were sacrificed and tumor masses were measured with a caliper (tumor mass in mm3 was calculated using the formula π/6 larger diameter x smaller diameter2) and analyzed by immunohistochemistry (IHC). Simultaneously, four groups of five 4–6-week-old mice animals totally irradiated (1.2 Gy,137Cesium source) were injected into the tail vein with 2 × 106 U266 cells. At day 7 post-injection, mice were daily treated with macitentan (30 mg/kg) or vehicle (Methocell 0,05% and TWEEN 80 0,05%) per os. Following a 7-day treatment, mice were sacrificed, and MM BM burden was evaluated as percentage of viable human CD45+CD138+ cells over total viable human CD45+ TOPRO3 negative cells. The day of beginning and the length of macitentan treatment were established based on preliminary experiments. RPMI-8226 and U266 cell lines were chosen on the basis of the literature data for subcutaneous and intra-tail injection of MM cell lines, respectively (22 (link)).
5
Leukemia Engraftment in NOG Mice
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NOD/Shi-scid/IL-2Rγnull (NOG) mice were purchased from Taconic (Germantown,
NY) and kept in pathogen-free conditions in the animal facility of the
Interdepartmental Centre of Experimental Research Service (CIRSAL) of the University
of Verona, as approved by the Italian Minister of Health. VR-ALL cells (5 ×
106) were injected via tail vein into 8 to 12 weeks old mice previously
irradiated with 1.2 Gy from a 137Cs source. Eight weeks following the
initial injection of VR-ALL cells, animals were sacrificed and leukemic burden was
quantified in organs as number of hCD45+ cells.
NY) and kept in pathogen-free conditions in the animal facility of the
Interdepartmental Centre of Experimental Research Service (CIRSAL) of the University
of Verona, as approved by the Italian Minister of Health. VR-ALL cells (5 ×
106) were injected via tail vein into 8 to 12 weeks old mice previously
irradiated with 1.2 Gy from a 137Cs source. Eight weeks following the
initial injection of VR-ALL cells, animals were sacrificed and leukemic burden was
quantified in organs as number of hCD45+ cells.
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