Nude mice (CAnN.Cg-Foxn1nu/CrlCrlj; female, 5–6 weeks old) were obtained from Charles River Laboratories Yokohama, Japan. Cancer cells (5 × 106 cells/head) were implanted s.c. When the tumor volume reached 100–300 mm3, mice were randomized into treatment groups. Lenvatinib or golvatinib were given orally once a day. The tumor volume was calculated by using the following formula: tumor volume (mm3) = 1/2 × length (mm) × width (mm)2. For the isolation of splenic macrophages, BALB/cAnNCrlCrlj mice (female, 5–6 weeks old) were obtained from Charles River Laboratories. All of the animal experiments were carried out in accordance with the guidelines for animal experiments of Eisai Co., Ltd.
Cann cg foxn1nu crlcrlj
Manufactured by Charles River Laboratories
Sourced in Japan
The CAnN.Cg-Foxn1nu/CrlCrlj is a laboratory mouse model. It is a nude mouse, meaning it lacks a functional thymus gland and exhibits a severe deficiency in T cell-mediated immune responses. This model is commonly used in research applications that require a compromised immune system.
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11 protocols using cann cg foxn1nu crlcrlj
1
In Vivo Tumor Xenograft Evaluation
2
Xenograft Model for Evaluating Anti-Cancer Agents
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Animal experiments were carried out in accordance with the Institutional Animal Care and Use Committee guidelines of Eisai Co., Ltd. Female BALB/c nude mice (CAnN.Cg‐Foxn1nu/CrlCrlj) were obtained from Charles River Laboratories Japan (Kanagawa, Japan). A‐498, Caki‐1, and Caki‐2 cells were suspended in 50% Matrigel (Corning, Corning, NY, USA) and KP‐1 transfectants were suspended in Geneticin‐free medium at 5 × 107–1 × 108 cells/mL; 0.1–0.2 mL cell suspension was inoculated s.c. into the right flank of each mouse. Once the tumors had reached the desired volume, the mice were selected according to tumor volume and shape and randomly allocated to treatment groups (day 1). Lenvatinib was dissolved in 3 mmol/L HCl or distilled water. Everolimus was dissolved in a solution comprising DMSO, Tween‐80, and 5% glucose (35:65:900 [or 1:2:27]), and PD173074 was dissolved in distilled water containing equimolar HCl. Each diluted drug or vehicle was given orally to individual mice once daily. Tumor dimensions were measured twice weekly with a caliper, and tumor volume was calculated as ½ × length × width2. The RTV was calculated by dividing the daily tumor volume by that of the same mouse on day 1. Body weight was also measured twice weekly, and the relative body weight was calculated by dividing the daily body weight by that of the same mouse on day 1.
3
Xenograft Model for ALK-driven Cancers
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Ba/F3 cells stably expressing DmrB-ALK_wt or DmrB-ALK_mF1174L, or Ba/F3 cells stably expressing EML4-ALK/EML4-ALK or EML4-ALK/EML4cc were subcutaneously injected into 8-w-old female BALB/C nu/nu mice (CAnN.Cg-Foxn1nu/CrlCrlj, Charles River Laboratories, Wilmington, MA, USA). For DmrB-ALK_wt or DmrB-ALK_mF1174L xenograft, subsequently, 0.5 mg/kg B/B solubilized in nano-pure water with 4% ethanol, 10% polyethylene glycol 400 (PEG-400), and 1.75% polyoxyethylene (20 (link)) sorbitan monolaurate (Tween-20), was injected twice a week into the intraperitoneal cavities of the mice. The control group received a mock solution (4% ethanol, 10% PEG-400, and 1.75% Tween-20 solubilized in nano-pure water). Tumor volumes were calculated twice a week with a modified ellipsoid formula: 1/2 × (width2 × length) for 5 w. In two of the six mice injected with Ba/F3 DmrB-ALK_wt, administration of B/B was discontinued after 3.5 w and changes in tumor volume were observed. The animal study protocol was approved by the Asahikawa Medical University Research Ethics Committee (#18133).
4
Mouse Hindlimb Ischemia Model and Cell Therapy
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Eight‐ to 10‐week‐old male BALB/c nu/nu mice (CAnN.Cg‐Foxn1nu/CrlCrlj; Charles River Laboratories Japan, Inc., Tokyo, Japan) were used, as reported elsewhere.26 (link) The proximal portion of the left femoral artery, including the superficial and the deep branch, was suture‐ligated, and the proximal and distal portions of the saphenous artery were occluded with a bipolar forcep electric coagulator (MERA N3‐14; SENKO MEDICAL INSTRUMENT mfg. Co., Ltd., Tokyo, Japan). The overlying skin was closed with a 6‐0 silk suture. The next day, cells were suspended in IMDM medium and intramuscularly injected into ischemic hindlimbs.
The cell injection sites and the doses for assays were as follows: each one site of anterior tibial muscle (ATM) and gastrocunemius muscle (GCM) for blood flow analysis and histology, that is, hematoxylin and eosin (H&E) staining, Azan staining, and inducible nitric oxide synthase (iNOS) immunohistochemistry (IHC) (5.0×103 cells/20 μL per site: total 1×104 cells/mouse), 2 sites of ATM for qRT‐PCR (5.0×103 cells/20 μL per site: total 1×104 cells/mouse), or for histological assessment by confocal images (1.0×105 cells/20 μL per site: total 2×105 cells/mouse).
The cell injection sites and the doses for assays were as follows: each one site of anterior tibial muscle (ATM) and gastrocunemius muscle (GCM) for blood flow analysis and histology, that is, hematoxylin and eosin (H&E) staining, Azan staining, and inducible nitric oxide synthase (iNOS) immunohistochemistry (IHC) (5.0×103 cells/20 μL per site: total 1×104 cells/mouse), 2 sites of ATM for qRT‐PCR (5.0×103 cells/20 μL per site: total 1×104 cells/mouse), or for histological assessment by confocal images (1.0×105 cells/20 μL per site: total 2×105 cells/mouse).
5
Eribulin Treatment on MX-1 Xenograft Tumors
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For in vivo xenograft specimens, 10 × 106 MX-1 cells in a Matrigel suspension were subcutaneously injected into the right flank of athymic mice (CAnN.Cg-Foxn1nu/CrlCrlj; Charles River Laboratories Japan, Yokohama, Japan). Nine to eleven days after inoculation, eribulin (0.3, 1, and 3 mg kg−1) or vehicle was intravenously administrated; this was defined as day 1. On days 4 or 8, mice were killed, and tumours were isolated and measured. Collected tumours were fixed in 10% neutral buffered formalin solution (Wako Pure Chemical Industries, Osaka, Japan) for 24 h followed by embedding in Tissue-Tek VIP 5 Jr. (Sakura, Torrance, CA, USA).
6
Xenograft Model of KTOR71 Cells
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Female BALB/c‐nu mice (CAnN.Cg‐Foxn1nu/CrlCrlj) were purchased at 6 weeks of age from Charles River Laboratories, Japan. To generate the xenograft model, KTOR71 cells (2 × 106) were suspended in Matrigel (Corning) and injected subcutaneously into the backs of the mice. The width and length of the tumors were measured every 3 days using electronic calipers. Tumor volume was calculated using the following formula: (length × width2) × 0.52. Treatment was initiated when the tumor volume reached 200 mm3. All animal experiments were approved by the Animal Research Committee of Kyoto University (ID: MedKyo 19,294, 20,256, and 21,272) and conducted in accordance with ARRIVE guidelines.
7
Xenograft Tumor Growth Assay
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Cells were admixed with 75 µL Matrigel (BD Biosciences) and the cell mixture was injected into both flanks of six-week-old NOD/SCID mice or nude mice. NOD/SCID mice (NOD.CB17-Prkdcscid/J) and nude mice (CAnN.Cg-Foxn1nu/CrlCrlj) were purchased from Charles River Laboratories Japan, Inc. (Yokohama, Japan). Tumors larger than 100 mm3 were counted. Tumor volume was measured two times a week using the following formula: V = 1/2(L × W2), where L equals length, and W equal width. Sample sizes were chosen to assure reproducibility of the experiments in accordance with the replacement, reduction, and refinement principles of animal ethics regulation. All animal studies were approved by the Kyoto University Animal Research Committee.
8
Xenograft Mouse Model for Anti-Cancer Therapy
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All animal experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals at Chugai Pharmaceutical Co., Ltd., and all animal procedures were reviewed and approved by the Institutional Animal Care and Use Committee at Chugai Pharmaceutical Co., Ltd. (approval number: 15-362). Five to seven-week-old male BALB/c-nu/nu mice (CAnN.Cg-Foxn1nu/CrlCrlj, purchased from Charles River Laboratories Japan) were inoculated subcutaneously in the right flank with 5 × 106 cells/mouse with either OE19b or OE19bTDR cells. Several weeks after the inoculation, mice were randomized to the control group or treatment groups. T-DM1 (10 mg/kg) or saline was administered intravenously once every 3 weeks. TRAS (40 or 80 mg/kg) and/or PER (40 or 80 mg/kg), or HuIgG was administered intraperitoneally once a week for 3 weeks. CAPE (359 mg/kg) or vehicle was administered orally for the initial 14 days. Tumor volume and body weight were measured twice a week. Tumor volume was calculated as described previously [8 (link)].
9
Xenograft Tumor Growth Inhibition
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Nude mice (CAnN.Cg-Foxn1nu/CrlCrlj; 5–6-week-old females) were obtained from Charles River Laboratories Japan (Kanagawa, Japan). Mice were maintained under specific pathogen-free conditions and housed in barrier facilities on a 12:12-h light:dark cycle, with food and water ad libitum. Cultured tumor cells (5 × 106 cells/mice) were implanted s.c. into the flanks of the mice. When the tumor volume reached 100–300 mm3, the mice were randomized into groups. Lenvatinib and golvatinib were dissolved in sterile distilled water and given orally once a day for 8–21 days. Mice were killed 24 h after the final treatment. Following treatment, each tumor was measured in two dimensions, and the volume was calculated by using the following formula: tumor volume (mm3) = 1/2 × length (mm) × [width (mm)]2. All animal experiments were carried out in accordance with the guidelines for animal experiments of Eisai Co., Ltd. Differences in relative tumor volume between vehicle-treated and compound-treated groups were analyzed using one-way anova followed by Dunnett's multiple-comparison test. Combination effects were analyzed using two-way anova . P-values of <0.05 (two-sided) were considered statistically significant.
10
Nude Mouse BALB/c Xenograft Model
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Female nude mice with BALB/c genetic backgrounds (CAnN.Cg-Foxn1nu/CrlCrlj, 5 weeks old) were purchased from Charles River Japan, Inc. (Kanagawa, Japan), maintained under specific pathogen-free conditions in a 12:12 h light-dark cycle, and provided with sterile food and water ad libitium. Animal studies were performed in accordance with the procedures approved by the animal use and care committee of the Japanese Foundation for Cancer Research.
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