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Nmri nu nu mice

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The NMRI nu/nu mice are a laboratory mouse strain that lack a functional thymus, resulting in a deficiency in T cells. These mice are often used in research applications that require an animal model with a compromised immune system.

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16 protocols using nmri nu nu mice

1

Preclinical cancer models for physical activity

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The experiments were performed in female athymic NCr nu/nu (age 7–10 weeks; Taconic Biosciences, NY), female NMRI nu/nu mice (age 30 weeks; Janvier Labs, France), male C3H mice (age 16 weeks; Janvier Labs) and male and female CDF1 mice (age 16 weeks and 24 weeks respectively; Janvier Labs). All mice were maintained in a pathogen-free facility and provided with water and food ad libitum. The mice strains used in the experiments are frequently used in oncology research. Specifically, athymic nude mice and CDF1 females represent models for breast cancer research [21 (link),22 (link)] and both CDF1 male and C3H male mice are used in pre-clinical colon cancer research [23 (link)]. Epidemiological data has shown that the risk of developing several cancers including both breast cancer and colorectal carcinoma risk can be reduced by physical activity [24 (link)].
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2

Therapeutic MSC and Bortezomib for Colo205 Tumors

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Animal care and all experiments performed were in accordance with federal guidelines and had been approved by university and state authorities. Female NMRI nu/nu mice (Janvier), 8 weeks old, were injected subcutaneously (s.c.) with 3 × 106 Colo205 cells in 100 μl PBS at left and right dorsal sides. Treatment started 10 days after tumor cell inoculation when tumors reached ~100 mm3. In particular, 4 × 106 MSCs were resuspended in 100 μl PBS mixed with 100 IU/ml of heparin (29 (link)) and then peritumorally injected (p.t.). During the injections of all cell lines, mice were anesthetized with isoflurane. The Colo205-bearing mice received three p.t. injections of MSCs at day 10, 17, and 27. In addition, 5 μg of BZB in 100 μl PBS were injected i.p. every second day, starting from day 11 until day 31. Mice in the control groups received either 100 μl PBS i.p. injected or MSCs.Mock. Tumor growth was monitored as described (30 (link)). Blood samples were taken from the tail at day 31, centrifuged (10,000 × g, 10 min, 4°C) and then stored at −80°C. Activity of ALT was determined by an enzymatic assay accordingly to the manufacturer’s instructions (BIOO Scientific, Austin, TX, USA).
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3

In Vivo Xenograft Experiments in Immunodepressed Mice

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Six- to eight-week-old immunodepressed female NMRI-nu/nu mice (JANVIER Labs, Le Genest-Saint-Isle, France) were used for in vivo experiments to limit immune response against NPC cells. All the experiments involving mice were done using appropriate conditions of husbandry experimentation and care, under the control of the Regional Comity of Midi-Pyrénées (France). For the guidelines on animal welfare, we followed the European directive 2010/63/EU. For administration of substances and removal of blood, including routes and volumes, we followed the good practice guide published in 2001 by Diehl et al.22 (link) The ethics committee, which approved the research, is CEEA-122, chaired by Dr Nicolas Cenac.
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4

Subcutaneous Xenograft Mouse Model

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All experiments were performed in accordance with animal care guidelines from the European Union and were approved by the appropriate authority. The animal project registered under the number (#2438) received a favorable assessment from the Ethics Committee and was approved by the French Higher Education and Research Minister. All procedures involving animals were performed under general anesthesia with inhaled isoflurane (Vetflurane; Virbac, France) using a Univentor 400 anesthesia unit (Genestil, Royaucourt, France). Mice were housed in filtered air cabinets with a 12 h light/dark cycle at 22–24 °C and 50% humidity, provided with food and water ad libitum, and manipulated following aseptic procedures. Female NMRInu/nu mice (Janvier, St Berthevin, France) aged 9–10 weeks were used, with a mean bodyweight of 30 ± 3 g. Mice were inoculated subcutaneously in the left flank with 8 × 106 exponentially growing HT29 cells and the experiments were initiated 5–7 days after inoculation when the tumors reached 50 mm3 in volume.
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5

Metformin and GANT61 Enhance Radiotherapy

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Animal experiments were approved by the ethics committee of KU Leuven (P131/2014). Male NMRI Nu/Nu mice (Janvier, France) were inoculated in both flanks with 2 × 106 22Rv1 cells in 100:100 µL medium/Matrigel (VWR, Radnor, PA, USA). Once tumors reached a volume of 150 mm3, mice were treated by intraperitoneal injection with solvent (9:1 saline/EtOH), metformin (250 mg/kg, every day), GANT61 (50 mg/kg every other day) or the combination of both drugs for 7 weeks. At day 5 of drug treatment, tumors were irradiated with a single dose of 6 Gy. During the entire treatment period, tumor growth was followed by 2-weekly caliper measurements and tumor volumes were calculated (V = (length × width × height) × π/6). In addition, the body weight of the mice was monitored to assess potential treatment toxicity. Mice were euthanized at the end of drug treatment or when tumors reached the maximum ethically permitted volume of 2 × 103 mm3. Thirty min before euthanasia, pimonidazole was intraperitoneally injected. Afterwards, tumors were excised and half of the tumor was fixed in formalin and embedded in paraffin for immunohistochemical analysis and the other half was snap-frozen for protein analysis.
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6

Tumorigenicity Assay of OVCAR-5 Cells

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Ovarian cancer cells OVCAR-5 were sorted for CXCR4 and CD133 as previously described29 (link). For tumorigenicity assays, serial dilutions of single-cells re-suspended in MatrigelTM (BD Bioscience, Heidelberg, Germany) were subcutaneously injected into female nude NMRI nu/nu mice (Janvier, Le Genest-Saint-Isle, France).
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7

Establishing Xenograft Models of Liposarcoma

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Female adult, partially immunodeficient, athymic NMRI nu/nu mice (JANVIER LABS, Saint Berthevin, France) were used for establishing xenograft models and for the in vivo experiments. Collection and usage of tumor samples from consenting patients were approved by the Medical Ethics Committee, University Hospitals Leuven. Animal experiments were approved by the Ethics Committee for Laboratory Animals, KU Leuven (Leuven, Belgium).
The SW872 liposarcoma cell line (Cell Lines Service, Eppelheim, Germany) was cultured in Dulbecco's modified Eagle's medium/F12 medium with 10% FBS (all from Life Technologies). The SW872 cell line has been previously studied in vitro and in vivo[15] (link), [16] (link). The SW872 model was generated by subcutaneous, bilateral injection of 5 × 106 cells per mouse site. Patient-derived DDLPS xenografts (UZLX-STS3 and UZLX-STS5) were established by bilateral subcutaneous implantation of fresh surgically resected tumor specimens from patients with DDLPS. Tumor tissue was further re-transplanted from mouse to mouse at least twice. From each passage, tumor fragments were collected for histologic and molecular characterization.
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8

Pharmacokinetics of Anti-Proliferative Agent

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All animal experiments
were conducted in accordance with the German Animal Welfare Law and
were approved by local authorities. Female NMRI nu/nu mice (Janvier,
France) were dosed once at 100 mg/kg orally (p.o., in PEG400/ethanol 9:1) (n = 3 per group), intraperitoneally
(i.p., in solutol/ethanol/water 4:1:5), or subcutaneously
(s.c., in castoroil/benzylbenzoate 9:1). After 0.5,
3, 6, 14, 24, and 48 h, mice were sacrificed by decapitation and blood
sampled in potassium-EDTA tubes (Sarstedt, Germany). 100 μL
of plasma was used for analysis. Samples were precipitated by the
immediate administration of ice cold acetonitrile in a 1:5 dilution.
Samples were frozen at −20 °C overnight and subsequently
centrifuged at 3000 rpm for 15 min. The supernatant was analyzed with
an Agilent 1200 HPLC system with MS/MS detection (AB Sciex, Framingham,
MA). Obtained exposures were corrected for plasma–protein binding
and reported in relation to the antiproliferative IC50u.
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9

Mouse Embryo Development Protocol

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All animal experiments were conducted according to the European and Belgian legislation and with approval of the Animal Ethics Committee of the faculty of Biomedical Sciences of the KU Leuven (project license 221/2015). SWISS, NMRI nu/nu mice (Janvier labs) and B6.Cg-Tg(ACTb-eGFP) mice were used in these studies. For timed matings, male and female SWISS mice were placed together and left overnight. The next day (0.5 dpc) the success of the mating was assessed by the presence of a vaginal plug. At 14.5 dpc pregnant females were sacrificed by cervical dislocation.
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10

Xenograft Engraftment in Nude Mice

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Nine NMRI nu/nu mice (Janvier Laboratories, Le Genest-St-Isle, France) aged 8 weeks were used as recipients for the xenografts. They were housed in cages under filtered hoods (MicroIsolator, Uno) in rooms maintained at an ambient temperature between 22 and 24°C with a day/night cycle of 12 h. All housing material and food were autoclaved before use. The mice were fed ad libitum on laboratory chow (complete food for rats and mice; Pavan Carfil) and acidified water. All experiments in this study were approved by the Ethics Review Board and the Committee on Animal Research of the Catholic University of Louvain.
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