All animal studies were performed in athymic nu/nu mice (Envigo) following Institutional Animal Care and Use Committee (IACUC) approval at the University of Minnesota. For subcutaneous tumor implantation, animals (n=3–4/group) received unilateral injections on the right shoulder of 1×106 CWR-R1 or CWR-R1CD133 cells (in 100 μL) in a 1:1 dilution of Matrigel (Corning) to 1x PBS. Tumors were measured twice weekly with calipers and volumes were calculated as length × width × height. For the intracardiac dissemination model, animals (n=4/group) received injections of 2×105 CWR-R1-EnzR or CWR-R1-EnzRCD133 cells (in 100 μL) in 1x PBS directly into the left ventricle of the heart, with a 75% accuracy rate. Following the intracardiac injections, mice were weighed twice/week to assess overall health and bioluminescent imaging was performed once/week to evaluate tumor formation and growth.
Athymic nu nu mice
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Athymic nu/nu mice are a strain of mice that lack a functional thymus gland, resulting in a deficiency of T cells. This mouse model is commonly used in biomedical research, particularly in the areas of cancer research and immunology.
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7 protocols using athymic nu nu mice
1
Investigating Prostate Cancer Metastasis
2
Xenograft Model for Neratinib and Alpelisib
3
Athymic Nude Mice Xenograft Model
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For this study, we used six-week-old female athymic nu/nu mice (Envigo) weighing 18–22 g. These animals were kept in sterile conditions, with autoclaved cages, bedding, water, and food, with 12 h of light and 12 h of dark cycle. Because of the patients’ young age, their tutors gave written consent for research purposes. The in vivo design of experiments was approved by the IDIBELL animal facility committee (AAALAC Unit1155) and the Institutional Ethics Committees approved the study protocol. All experiments followed the Ethical Conduct in the Care and Use of Animals guideline as indicated by The International Guiding Principles for Biomedical Research Involving Animals, developed by the Council for International Organizations of Medical Sciences.
4
Orthotopic Pancreatic Tumor Xenograft
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Female six week old athymic-nu/nu mice were purchased from Envigo and handled in accordance with the University of Iowa Animal Care and Use Committee (IACUC). MIA PaCa-2 cells (4 × 105) expressing luciferase and GFP were injected orthotopically directly into the pancreas of mice using ultrasound guidance as previously described [24 (link)]. Cells were suspended in a 1:1 mixture of PBS and Matrigel (Corning, St. Louis, MO) immediately prior to injections to prevent cells from dispersing throughout the abdomen. Tumor formation and growth were monitored weekly through bioluminescent imaging. Mice were given IP injections (200 µl) of a 15 mg/ml solution of VivoGloTM luciferin (Promega, Madison, WI). Following injection, mice were anesthetized by isoflurane inhalation and imaged using the AMI-1000 (Spectral Instruments Imaging). Total photon flux (photons per second) was quantified (AMIView software) by placing and area of interest around each mouse. Tumor progression was allowed to continue for fifty days post cancer cell injection.
5
Athymic Mouse Study on Augusta University
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All animal studies were approved
by the Institutional
Animal Care and Use Committee at Augusta University. Athymic nu/nu
mice were purchased from Envigo. All animal procedures and maintenance
were conducted in accordance with the institution approved guidelines
of Augusta University.
by the Institutional
Animal Care and Use Committee at Augusta University. Athymic nu/nu
mice were purchased from Envigo. All animal procedures and maintenance
were conducted in accordance with the institution approved guidelines
of Augusta University.
6
Preclinical Evaluation of Antibody-Drug Conjugates
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All animal experiments were IACUC approved and performed according to AAALAC guidelines and NIH best practices in AAALAC-accredited facilities at Vanderbilt University. MDA-MB-231, MDA-MB-231.Luc, HCC70, or HCC1187 cells (1 × 106) in 100 μL of 50% Matrigel were injected into the inguinal mammary fat pads of 4–6-week-old female athymic (nu/nu) mice (Envigo). Mice were randomized into treatment groups when tumor volume reached 50 mm3 as measured by calipers using the formula Tvol = (length X width2) / 2. Mice were treated by intravenous (i.v.) delivery at the indicated doses, durations, and frequencies. Conjugates were delivered in 0.9% saline. MIK665 treatments were delivered in 2% vitamin E/d-α-tocopheryl polyethylene glycol 1000 succinate (Sigma-Aldrich) in NaCl 0.9% (wt/vol) and delivered i.v. via tail-vein at 12.5 mg/kg at the indicated schedule. Mice were humanely euthanized at treatment day 4, 8, 28, or 35 as indicated, or when tumors exceeded 1000 mm3. Tissues and tumors were collected at necropsy, and frozen or processed for analysis.
7
Angiogenesis Quantification in Xenograft Model
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