All animals were maintained in accordance with IACUC standards of care in pathogen-free rooms, in the Moffitt Cancer Center and Research Institute (Tampa, FL) Vivarium. One week before inoculation with tumor cells (5 × 106 OVCAR3 cells, subcutaneously), 6–8 weeks old, female NSG mice (Charles River Laboratories) were assigned to one of the following four treatment arms: 1: Control group, treated with vehicle (DMSO) intraperitoneally. 2: MTD group, treated with PARPi (Olaparib), 100 mg/kg intraperitoneally, three times per week. 3: AT1 group, which was treated with PARPi (Olaparib) by the AT1 algorithm (dose modulation; see below). 3: AT2 group, which was treated with PARPi (Olaparib) by the AT2 algorithm (dose skipping; see below). Tumor growth was monitored every other day and tumor size was measured by calipers three times a week (Monday, Wednesday, Friday). These measurements were used to inform the dose choices under AT1 and AT2 at these times. Tumor volume was calculated using the following formula: volume = π (short diameter)2 × (long diameter)/6. When the tumor volume reached 200 mm3, treatment was started. Animal weights were measured and recorded twice weekly, and the overall health of each animal was noted to ensure timely end points within the experiment. Finally, animals were humanely killed and tumors were extracted.
Female nsg mice
Manufactured by Charles River Laboratories
Sourced in Canada
Female NSG mice are a specific strain of immunodeficient mice used in biomedical research. They are genetically engineered to lack mature T cells, B cells, and natural killer cells, making them useful for the study of human diseases and the evaluation of human cell and tissue xenografts.
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Lab products found in correlation
Vernier caliper, by Mitutoyo (1 mentions)
Female balb c mice, by Inotiv (1 mentions)
Accumax, by Innovative Cell Technologies (1 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Recombinant human il 2, by Miltenyi Biotec (1 mentions)
Hanks balanced salt solution (hbss), by Euroclone (1 mentions)
Matrigel, by BD (1 mentions)
11 protocols using female nsg mice
1
PARP Inhibitor Dose Optimization in Ovarian Cancer
2
Mammary Fat Pad Tumor Growth Assay
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14 Female NSG mice were purchased from Charles River Laboratories (Sain-Constant, QC, Canada), housed and maintained under specific pathogen-free conditions (RI-MUHC animal facility). The mice were randomly divided into two groups (n = 7 mice per group). At 7 to 9 weeks of age, the first group was injected in the fourth-left mammary fat pad with 1 × 106 MDA-MB-231-Scr and the second group was injected with MDA-MB-231-Sh-KIF5B. Tumor growth were monitored up to 7 weeks after injection. When tumors were detectable, tumor size was measured with a vernier caliper (Mitutoyo, Kawasaki, Japan) and calculated using the formula [length + width2]/2. Mice were sacrificed by CO2 asphyxiation.
3
Intratracheal Tumor Xenograft Model
4
Targeted Nanocarrier-Mediated Tumor Imaging
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Female NSG mice (9 weeks old, Charles River) were challenged by s.c. injection of A375 (1 × 106) cells on the right flank with a 23-gauge needle. Amphoteric ICG-NCs and cationic ICG-NCs were administered i.v. at day 14 post-tumor challenge at 35 mg/mL of NCs (160 μL, 50 μM of ICG). PBS and ICG free were injected to control groups. Fluorescence was monitoring by in vivo imaging system (IVIS; Perkin Elmer, Waltham, MA, USA) using excitation/emission wavelengths of 780 nm/845 nm.
5
Orthotopic Lung Tumor Xenograft Model
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1 × 106 NCI-H460 FLuc cells were administered by a non-invasive intratracheal technique into the lungs of female NSG mice aged 6-9 weeks (Charles River Laboratories) based on previously described methods 41 (link), 42 (link). Mice were anaesthetized with isoflurane (2-2.5% in O2) and transferred onto a vertical board where they were suspended in an upright position by their upper incisors. Using a Leica M125 stereomicroscope (Leica Microsystems), the tongue was moved to one side to expose the vocal cords and the entrance to the trachea, where a plastic 20 G i.v. catheter, attached to a 1 mL syringe was inserted. Approximately 50 µL of air was first injected into the lung to inflate it prior to the administration of 20 µL of cell suspension in PBS. Tumor growth was monitored through bioluminescent imaging. For further details see Supplemental Methods, Figures and Tables .
6
Subcutaneous Tumor Implantation and Imaging
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All in vivo experiments were conducted under the authority of project and personal license granted by the UK Home Office and the UKCCCR Guidelines (1998). Female Balb/c mice aged 4-6 weeks (Envigo, UK) were inoculated subcutaneously with CT26 cells (1 × 106 cells in 0.1 mL PBS) in the lower flanks. Female NSG mice aged 4-6 weeks (Charles River, UK) were inoculated subcutaneously with PANC-1 cells (5 × 106 cells in 0.1 mL PBS) in the lower flanks. Nuclear imaging studies were conducted in mice carrying one tumor in one flank; biodistribution and therapy studies were performed in mice carrying two tumors one in each flank. Normal C57/Bl6 mice aged 4-6 weeks (Envigo, UK) without tumor inoculation were used for the assessment of the biodistribution of THP-amine.
7
Subcutaneous Cell Injection in NSG Mice
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Cells were suspended in PBS, and 100 μL of the cell mixture was injected subcutaneously into 8-week-old female NSG mice (Charles River). No blind experiments were done.
8
In vivo Tumor Growth and Metastasis
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For tumor growth, 1 × 105 SPANXB1 and empty vector transduced MCF-7cells in 1:1 mixture of PBS and matrigel were injected in the mammary fat pad of 4–6 week old, female NSG mice (Charles River)27 (link). All experiments were performed in accordance with the Animal Care and Use Committee guidelines. Each group consisted of 5 mice. Mice were examined every day and mice showing any sign of morbidity were immediately sacrificed according to the University guidelines. All experiments were terminated at week 5 due to the tumor burden associated effects. After 5 weeks, mice were sacrificed and tumor weights were taken. Lungs and livers were removed for the analysis of metastasis. Focal tumor nodules were counted in the lung and liver of all the mice from various groups. Tumors were immediately snap frozen and also processed as formalin fixed paraffin embedded tissues (FFPE) for histological, Western blotting and IHC analyses respectively. All histopathological evaluations of the in vivo tumors were done per pathologic guidance27 (link). Data presented as mean ± SE of duplicate experiments.
9
Breast Cancer Metastasis Model in NSG Mice
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12 Female NSG mice were purchased from Charles River Laboratories (Sain-Constant, QC, Canada), housed and maintained under specific pathogen-free conditions (RI-MUHC animal facility). The mice were randomly divided into two groups (n = 6 mice per group). At 7 to 9 weeks of age, the first group was injected in the tail vein with 5 × 105 T47D-Si-control and the second group was injected with T47D-Si-KLC1. Mice were monitored up to 5 weeks after injection. Mice were sacrificed by CO2 asphyxiation and lungs were collected.
10
Subcutaneous Organoid Xenograft Assay
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Organoids were dissociated by TrypLE (ThermoFisher) or Accumax (Innovative Cell Technologies). 1 × 105 cells/100 μl of 50% Matrigel in DMEM/F12 were transplanted subcutaneously to NSG female mice (Charles River) as described previously44 (link). Four weeks later, mice were euthanized, and subcutaneous tumors were collected, measured, and processed with HE staining. The animal experimental protocol defines that subcutaneous tumor size should not exceed 10% of the mouse body weight. In our experiment, the tumor size was not exceeded.
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