Male nude mice (BALB/c) at 8 weeks of age were obtained from Japan SLC. The treatment protocol followed the guidelines for animal experimentation adopted by Mie University, and meets the standards required by the UKCCCR guidelines.19 (link) To establish a mouse peritoneal metastasis model, NUGC3 gastric cancer cells (3 × 106 cells/ml/mouse) transfected with L1CAM siRNA or negative control siRNA were injected intraperitoneally into mice under Isoflurane inhalation (Mylan, Tokyo, Japan), as previously described.14 (link) Further information was described in Supplementary file .
Male balb c nude mice
Manufactured by Japan SLC
Sourced in Japan
Male BALB/c nude mice are an inbred strain of laboratory mice that lack a functional thymus gland, resulting in a severe deficiency of T cells. They are commonly used in biomedical research, particularly in the fields of oncology and immunology, where their immunodeficient nature allows for the study of human tumor xenografts and the evaluation of novel therapeutic approaches.
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Lab products found in correlation
T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Matrigel, by BD (1 mentions)
L glutamine, by Nacalai Tesque (1 mentions)
Eagle s minimum essential medium, by Fujifilm (1 mentions)
Stromal cell basal medium, by Lonza (1 mentions)
Penicillin streptomycin, by Nacalai Tesque (1 mentions)
Rpmi 1640 medium, by Nacalai Tesque (1 mentions)
5 protocols using male balb c nude mice
1
Establishment of Peritoneal Metastasis Model
2
Antitumor Effect of K252a in Peritoneal Metastasis
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Male nude mice (BALB/c) at 8 weeks of age were purchased from Japan SLC Inc. (Shizuoka, Japan). The experimental protocols were reviewed and approved by the Animal Care and Use Committee at the Mie University Graduate School of Medicine. DLD1 cells (5×107 cells/500 µl PBS) were injected intraperitoneally for peritoneal metastatic formation. Either K252a (500 µg/kg) or PBS (control) was injected intraperitoneally three times a week to examine the effect of K252a on the peritoneal metastatic formation. Four weeks later, the mice were sacrificed, and then the size and number of peritoneal metastatic nodules was evaluated (each group; n = 5). Results are presented as mean ±SE.
3
Sorafenib Inhibits Tumor Growth in BALB/c Nude Mice
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Male BALB/c nude mice (5–6 weeks old) were purchased from Japan SLC, Inc. (Shizuoka, Japan) and acclimated to our facilities for one week. In the cTCC subcutaneous model, 2 × 106 Sora cells were subcutaneously implanted into the left flank of each mouse. Two days later, mice were randomly divided into two groups and treated with PBS or sorafenib (30 mg/kg) daily for 14 days. Subcutaneous tumor growth was recorded as the length and width of tumors using a Vernier caliper every 2 days, and the tumor size was calculated using the formula, (length × width × height)/2. On day 16, the mice were euthanized, and the tumors were collected for histological analysis and immunofluorescence. All procedures were approved by the Animal Care and Use Committee of The University of Tokyo (authorization number: P20-116).
4
In vivo Evaluation of Anticancer Drugs in Nude Mouse Xenograft Models
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Male BALB/c nude mice (five-week-old) kept in specific pathogen-free (SPF) conditions were from Japan SLC, Inc. (Shizuoka, Japan). The mice experimentation was conducted in compliance with the Animal Care and Use Committee, Okayama University.
T24/DOX cells (1×106) or T24/CIS cells (1×106) in 100 µl resuspension, mixing Matrigel (BD Biosciences, USA) and Hank's buffered saline solution (HBSS) at 1:1, were used for subcutaneous inoculation to mouse right flank. The mice bearing about 100 mm3 T24/DOX tumor received two-week oral administration of NTX (40 mg/kg) or PBS (n = 6 per group) once per day. It was the same with T24/CIS tumor-bearing mice. Both mouse body weight and tumor growth were measured twice a week. Surgical resection, weighing, and photography of T24/DOX and T24/CIS tumors were conducted at the indicated endpoint. The formula of volume = 0.52 × length × width2, was applied to calculate tumor growth. T-PER Tissue Protein Extraction Reagent (#78510, ThermoFisher Scientific, USA) was used for extracting tumor tissue protein, which was subsequently analyzed by western blot. For histological examination, T24/DOX and T24/CIS tumors underwent 10% formaldehyde fixation, paraffin imbedding, and 4 μm section cut. Hematoxylin and eosin (H&E) were applied to stain the sections, followed by observing NTX-induced morphological alterations.
T24/DOX cells (1×106) or T24/CIS cells (1×106) in 100 µl resuspension, mixing Matrigel (BD Biosciences, USA) and Hank's buffered saline solution (HBSS) at 1:1, were used for subcutaneous inoculation to mouse right flank. The mice bearing about 100 mm3 T24/DOX tumor received two-week oral administration of NTX (40 mg/kg) or PBS (n = 6 per group) once per day. It was the same with T24/CIS tumor-bearing mice. Both mouse body weight and tumor growth were measured twice a week. Surgical resection, weighing, and photography of T24/DOX and T24/CIS tumors were conducted at the indicated endpoint. The formula of volume = 0.52 × length × width2, was applied to calculate tumor growth. T-PER Tissue Protein Extraction Reagent (#78510, ThermoFisher Scientific, USA) was used for extracting tumor tissue protein, which was subsequently analyzed by western blot. For histological examination, T24/DOX and T24/CIS tumors underwent 10% formaldehyde fixation, paraffin imbedding, and 4 μm section cut. Hematoxylin and eosin (H&E) were applied to stain the sections, followed by observing NTX-induced morphological alterations.
5
Cell Culture and Animal Handling
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LNCaP cells, MCF7 cells, MDA-MB-231 cells, Caco-2 cells, HEK293 cells, and Cos7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Nacalai Tesque, Kyoto, Japan); PC3 cells were cultured in RPMI 1640 medium (Nacalai Tesque); and A375 cells were cultured in Eagle’s Minimum Essential Medium (Wako Pure Chemical Industries, Osaka, Japan). All media were supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Saint Louis, MO, USA), 20 mM l -glutamine (Nacalai Tesque) and 100 µ/mL penicillin–streptomycin (Nacalai Tesque). PrS cells were purchased from Lonza (Cat. No. CC-2508; Basel, Switzerland) and cultured in Stromal cell basal medium supplemented with growth factors (Lonza). Male BALB/c nude mice (7-weeks-old) were purchased from Japan SLC (Shizuoka, Japan). The animal experiments conducted in this study were approved by the Shiga University of Medical Science Animal Care and Use Committee according to the Animal Research Reporting of In Vivo Experiments guidelines..
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