To assay tumor formation, cells were harvested and resuspended in cold PBS. For A2780 xenograft experiments, 3 million cells were injected in each flank of NU/J mice (Jackson Laboratory, cat. no. 002019). For A2058 xenograft experiments, 2 million cells were injected in each flank in NU/J mice. For HCT116 xenograft experiments, 4 million cells were injected in J:NU mice (Jackson Laboratory, cat. no. 007850). For MCF10A cells expressing HRASG12V, 10 million cells were injected in each flank in NU/J mice. For AGS xenograft experiments, the following conditions were tried: 5 million cells resuspended in PBS in each flank of NU/J mice (Jackson Laboratory, cat. no. 002019); 4 million cells resuspended in PBS in each flank of NGS mice (Jackson Laboratory, cat. no. 005557); and 15 million cells resuspended in a 1:1 PBS:Matrigel mixture in each flank of J:NU mice (Jackson Laboratory, cat. no. 007850). Cells were subcutaneously injected using a 1 mL 25G x 5/8 syringe (BD, cat. no. 309626). Mice were visually monitored for tumor formation routinely following injection. Once a tumor was visible, it was measured every three days by calipers. Tumor volume was calculated using the formula V = ½ (longer axis)(shorter axis). All mouse protocols were approved by the CSHL and Yale Institutional Animal Care and Use Committees.
Nu j mice
Manufactured by Jackson ImmunoResearch
Sourced in United States, Montenegro, Germany
Nu/J mice are a specific mouse model developed by Jackson ImmunoResearch. They are characterized by a spontaneous mutation that results in a lack of functional T cells and B cells, leading to a severe combined immunodeficiency (SCID) phenotype. This mouse model is widely used in research to study the role of the immune system and for the development and testing of therapeutic interventions.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (6 mentions)
Matrigel, by Corning (3 mentions)
Balb c mice, by Jackson ImmunoResearch (2 mentions)
Ivis spectrum, by PerkinElmer (2 mentions)
Wild type c57bl 6j mice, by Jackson ImmunoResearch (2 mentions)
Vivoglo luciferin, by Promega (2 mentions)
C57bl 6, by Jackson ImmunoResearch (2 mentions)
Molm 14 cells, by Leibniz Institute DSMZ (2 mentions)
25 g x 5 8 syringe, by BD (1 mentions)
Syringe pump, by KD Scientific (1 mentions)
Purelink expi endotoxin free maxi plasmid purification kit, by Thermo Fisher Scientific (1 mentions)
Stereotaxic frame, by RWD Life Science (1 mentions)
Insulin, by Merck Group (1 mentions)
Mycoalert, by Lonza (1 mentions)
Gentlemacs tumor dissociation kit, by Miltenyi Biotec (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
Living image software, by PerkinElmer (1 mentions)
Anti asialo gm1, by Fujifilm (1 mentions)
Ab 105 c, by R&D Systems (1 mentions)
Adult sjl j mice, by Jackson ImmunoResearch (1 mentions)
66 protocols using nu j mice
1
Xenograft Tumor Formation Assay
2
Targeting PKCδ in Breast Cancer Stem Cells
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These studies were performed with the approval of the Institutional Animal Care and Use Committee of Boston University. Breast cancer stem cells (2 × 105) grown from a metastatic tumor were suspended in human breast cancer stem cell complete growth media (Celprogen, San Pedro, CA) and injected subcutaneous into the right flank of female J:NU mice (The Jackson Laboratory, ME) under anesthesia. After palpable tumors developed, the mice were divided into two groups of animals. The control group received daily intraperitoneal injections of vehicle (DMSO) while the treatment group received daily intraperitoneal injections of a PKCδ inhibitor (rottlerin 5,000 μg/kg) for 15 days. The length and width of tumors were measured with a vernier caliper and tumor volumes were calculated. Survival was calculated as the day tumors reached the maximum size allowed by the protocol (2 cm diameter).
3
Omega-3 PUFA Impacts Breast Cancer
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All animal procedures were performed in compliance with the guidelines approved by Massachusetts General Hospital (MGH) (IACUC). Male athymic nude (J:NU) mice purchased from Jackson Laboratory (Bar Harbor, Maine) were housed in SPF area in animal facility, MGH and were fed irradiated chow diet and autoclaved DI water ad libitum. The mice were randomly assigned to two groups, Control (n = 3) and n‐3 PUFA (n = 4), receiving 100 μL of corn oil (control) and omega‐3 oil (containing 90% of EPA + DHA) through oral gavage, respectively. Total duration of oil intervention included 30 days before and 45 days after tumor implantation. Prior to xenograft induction, MCF‐7‐CSC were grown in vitro in the presence of mammosphere‐formation medium described in method of cell culture. A cell suspension of 1.5 × 107 MCF‐7‐CSC in 150 μL of PBS was subcutaneously injected into the lower right flank. Following the xenograft induction, the mice were monitored for tumor growth and tumor size was measured using digital caliper in longest and shortest dimension. Formula = (longest length × shorted length2)/2 was applied to calculate the tumor size.
4
Establishment of Xenograft and Homograft Tumor Models
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All of our procedures were approved by the Louisiana State University Health Sciences Center Institutional Animal Care and Use Committee (IACUC), which is OLAW approved (Assurance #D16-00058), AAALAC accredited (#000037), and US Dept. of Agriculture certified (#72-R-0003). For MDA-MB-231 cell xenografts, 6-week-old, 15–20 g, female J/Nu mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA) and housed in a specific pathogen-free, climate-controlled colony room with a 12/12 h light/dark cycle with free access to water and lab chow. To establish xenografts, approximately 4 million cells were injected in a 50% matrigel/saline suspension. Tumors formed to 0.7–1.2 cm diameter in 3–4 weeks.
For 4T1 homografts, 6-week-old, 15–20 g, female BALB/c mice were purchased from Jackson Laboratories and housed as with the J/Nu mice, but in a room with modified barrier controls. To establish homografts, approximately 500,000 cells were injected subcutaneously between the scapulae in a saline suspension. Tumors formed to 0.7 to 1.2 cm in 7 days. Treatment day 1 was initiated when average tumor size attained 5–10 mm in length as measured with a digital caliper by at least 2 investigators, one of which was not blinded to the treatment groups.
For 4T1 homografts, 6-week-old, 15–20 g, female BALB/c mice were purchased from Jackson Laboratories and housed as with the J/Nu mice, but in a room with modified barrier controls. To establish homografts, approximately 500,000 cells were injected subcutaneously between the scapulae in a saline suspension. Tumors formed to 0.7 to 1.2 cm in 7 days. Treatment day 1 was initiated when average tumor size attained 5–10 mm in length as measured with a digital caliper by at least 2 investigators, one of which was not blinded to the treatment groups.
5
In Vivo Bioluminescence Imaging of AAV
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For AAV packaging, pAAV-CMV-AkaLuc-P2A-mNeonGreen-WPRE plasmid and pAAV-CMV-Antares-P2A-mNeonGreen-WPRE were amplified and purified by a PureLink Expi endotoxin-free maxi plasmid purification kit (Invitrogen) and were sent to Stanford Gene Vector and Virus Core to produce and titer AAV.DJ vectors. Under sterile conditions, 12-week-old male J:NU mice (Jackson Laboratory, 007850) were anesthetized with isoflurane and secured in a stereotaxic frame (RWD Life Science), and a hole the size of the needle was drilled through the skull. A Hamilton syringe with a 33-gauge needle was inserted at anteroposterior −2.2 mm, mediolateral +1.7 mm and dorsoventral −1.7 mm. After a 2-min wait, the needle was pulled back 0.3 mm, and 1 µl containing 1.4 × 1010 genome copies each of AkaLuc- and Antares-expressing AAV was injected at a speed of 0.1 μl min–1 using a syringe pump (KD Scientific). The needle was left in place for 3 min after each injection to minimize upward flow of viral solution. Two to 3 weeks after AAV infection, mice were injected i.p. with the indicated amount of CFz, Akalumine or FFz and imaged on the Ami HT (binning of 1 × 1 and exposure time of 10 s).
6
Xenograft Tumor Growth Modeling
7
Culturing Human Cancer and Fibroblast Cells
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The human glioblastoma U87 cell line and mouse 3T3 fibroblast cells were obtained from ATCC and cultured per the vendor's instructions. The human breast cancer MCF‐7 parental cell line, the P‐gp‐overexpressing MCF‐7 TX400 subline, and the ABCG2‐overexpressing MCF‐7 MX100 subline were cultured in EMEM supplemented with 10% FBS, 100 U mL−1 penicillin, 100 µg mL−1 streptomycin, and 0.01 mg ml−1 insulin (Sigma) as previously described.[18 (link)
] Cells were maintained in 5% CO2 at 37 °C and tested to be free of mycoplasma (MycoAlert, Lonza).
GBM39 cells expressing luciferase were serially passaged in the flank of female J:NU mice (4‐5 weeks old, #0 0 7850, Jackson Laboratory). Tumors were dissociated using a gentleMACS tumor dissociation kit (Miltenyi Biotec) before in vitro culturing. Cells were cultured as recommended by the Mayo Clinic Brain Tumor Patient‐Derived Xenograft National Resource using the FBS cell culture protocol. Briefly, cells were cultured in DMEM containing 2.5% FBS, 100 U mL−1 penicillin, and 100 µg mL−1 streptomycin for 48 h, then media was exchanged to DMEM containing 10% FBS and antibiotics for at least 24 h before experimentation. Cells were dissociated and used in experiments, but not passaged serially. Assays were performed within 14 days of tumor dissociation.
] Cells were maintained in 5% CO2 at 37 °C and tested to be free of mycoplasma (MycoAlert, Lonza).
GBM39 cells expressing luciferase were serially passaged in the flank of female J:NU mice (4‐5 weeks old, #0 0 7850, Jackson Laboratory). Tumors were dissociated using a gentleMACS tumor dissociation kit (Miltenyi Biotec) before in vitro culturing. Cells were cultured as recommended by the Mayo Clinic Brain Tumor Patient‐Derived Xenograft National Resource using the FBS cell culture protocol. Briefly, cells were cultured in DMEM containing 2.5% FBS, 100 U mL−1 penicillin, and 100 µg mL−1 streptomycin for 48 h, then media was exchanged to DMEM containing 10% FBS and antibiotics for at least 24 h before experimentation. Cells were dissociated and used in experiments, but not passaged serially. Assays were performed within 14 days of tumor dissociation.
8
Evaluating Novel Therapies for Pancreatic Cancer
9
Murine Model for Preclinical Research
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All experiments were conducted following NIH guidelines and protocols approved by the IACUC at Columbia University Irving Medical Center. 7–8-week-old female J:NU mice (The Jackson Laboratory) were housed in a barrier facility with ad libitum access to food pellets and water on a 12-h light-dark cycle. Mice were regularly monitored to follow institutional guidelines for ethical endpoints. All experimental endpoints were at 14–16 weeks of age unless otherwise specified.
10
Xenograft Tumor Models: EGFR and YAP Inhibitors
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Four-weeks old female nude (NU/J) mice were purchased from Jackson Laboratory and were six-weeks old at time of experiment. For subcutaneous xenotransplantation, H1975 (1×106 cells/flank) and HCC1359 (5×106 cells/flank) cells were re-suspended in a mixture of 50% PBS/50% Matrigel matrix and injected into the right and left flanks of mice. When the average tumor volume of H1975 reached 75 mm3, the mice were randomized into 2 groups to receive the following treatments: (a) Vehicle (2% DMSO, 20% PEG300, 2% Tween-80, 76% PBS); (b) Osimertinib (5 mg/kg, daily, p.o). The CIC KO HCC1359 tumor-bearing mice were also randomized into 2 groups: (a) Vehicle (2% DMSO, 20% PEG300, 2% Tween-80, 76% PBS); (b) Verteporfin (40 mg/kg, every 2 days, i.p). Tumor volume and body weight were measured every 2 days. Tumor volume was determined using caliper measurements of tumor length (L) and width (W) according to the formula V = (L × W2) × 0.52. Mice were observed post-procedure for 1–2 hours, and body weight and wound healing were monitored per IACUC protocol.
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