Male BALB/c‐nu mice (n = 16; 4‐5 weeks of age, 17‐21 g) were obtained from Vital River Laboratory Animal Technology, Beijing, China. The animal experiments were approved by the Committee on Ethics of Animal Experiments of Renmin Hospital of Wuhan University and performed in compliance with the Guide for the Care and Use of Laboratory Animals from the National Institutes of Health. The mice were reared under specific pathogen‐free conditions, with a 12 hour light/dark cycle and access to water and food ad libitum. The nude mice were then divided into two groups comprising the control group and the koPLAC8 group (both n = 8). Then, the cells (1 × 106; control or koPLAC8) were subcutaneously injected into the flank of each mouse. After 3 weeks, all the mice were killed. The xenograft tissue was dissected, fixed and stained.22
Male balb c nu mice
Manufactured by Charles River Laboratories
Sourced in China
Male BALB/c-nu mice are a well-established mouse model that exhibits a lack of thymus-dependent (T cell) immune function. These mice are useful for a variety of research applications that require an immunodeficient animal model.
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6 protocols using male balb c nu mice
1
Xenograft Model of Nude Mice
2
Celastrol inhibits colorectal cancer
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Human colorectal adenocarcinoma cell lines HCT116 and HT-29 were obtained from Shanghai Institute of Cell Resource Center of Life Science (Shanghai, China). All cells were cultured in McCOY’s 5A medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone, USA), 100 mg/ml streptomycin and 100 U/ml penicillin at 37°C in humidified atmosphere with 5% CO2.
Male C57BL/6 mice (n = 65, 6–8 weeks old) used for UC-CRC models and male BALB/c-nu mice (n = 36, 5 weeks old) used for colorectal tumor xenograft models were purchased from Vital River Laboratory Animal Technology Co. Ltd (Beijing, China). All animals were housed under controlled conditions (temperature 22 ± 1°C, humidity 40–60% and 12 h dark/light cycle) and free access to a standard laboratory diet and water for 2 weeks. All animal care and experimental procedures were carried out in accordance with the recommendation of the Animal Care Ethics and Use Committee of China Medical University and approved by this Committee.
Celastrol (≥98%) was purchased from Dalian Melone pharmaceutical Co., Ltd (Dalian, China). For in vitro studies, Celastrol was dissolved in dimethyl sulfoxide (DMSO; Sigma–Aldrich) at a stock concentration of 44 mM. For animal experiments, Celastrol was dissolved in DMSO at 20 mg/ml and then diluted with 0.9% saline to the final concentrations (1% DMSO) before administration.
Male C57BL/6 mice (n = 65, 6–8 weeks old) used for UC-CRC models and male BALB/c-nu mice (n = 36, 5 weeks old) used for colorectal tumor xenograft models were purchased from Vital River Laboratory Animal Technology Co. Ltd (Beijing, China). All animals were housed under controlled conditions (temperature 22 ± 1°C, humidity 40–60% and 12 h dark/light cycle) and free access to a standard laboratory diet and water for 2 weeks. All animal care and experimental procedures were carried out in accordance with the recommendation of the Animal Care Ethics and Use Committee of China Medical University and approved by this Committee.
Celastrol (≥98%) was purchased from Dalian Melone pharmaceutical Co., Ltd (Dalian, China). For in vitro studies, Celastrol was dissolved in dimethyl sulfoxide (DMSO; Sigma–Aldrich) at a stock concentration of 44 mM. For animal experiments, Celastrol was dissolved in DMSO at 20 mg/ml and then diluted with 0.9% saline to the final concentrations (1% DMSO) before administration.
3
Xenograft Tumor Model in Mice
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Male BALB/c-nu mice aged 6 weeks old and weighing approximately 15 g each were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were kept at temperature (22-24°C) with a stable humidity (55±15%) with free access to food/water in a 12 h/12 h light/dark cycle and housed in specific pathogen-free conditions at Jilin University (19 (link)). A total of 5×106 SNU-449 cells were subcutaneously injected into the backs of the mice. After 7 days, when the tumor volume reached approximately 100 mm3, the nude mice were randomly divided into 4 groups and were administered treatment intraperitoneally (i.p.) every day (Fig. 3A ). The control group, administered 200 µl saline; the DIC group, administered 25 mg/kg DIC; the OXA group, administered 200 mg/kg OXA (Shandong New Time Pharmaceutical Co., Ltd.); and the OXA + DIC group, administered the corresponding drugs indicated above. All animal experiments were performed in accordance with the National Guidelines for Experimental Animal Welfare and with approval of the Animal Welfare and Research Ethics Committee at Jilin University (Changchun, China).
4
Xenograft and Pulmonary Metastasis Mouse Models
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Male BALB/c-nu mice (4-weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China), and maintained in laboratory animal facility of Zhongnan Hospital of Wuhan University. After adaptive feeding for a week, xenograft models were established by subcutaneously inoculating 1 × 106 UM-UC-3 LV-NC cells or LV-CIRBP sh cells diluted in100 µl PBS (n = 4), while pulmonary metastasis models were established by tail intravenous injecting 1 × 106 cells diluted in 100 µl PBS (n = 3). 6 weeks post injection, the mice were sacrificed and the tumors were taken out and weighed, meanwhile the tumor volume was measured every 6 days (tumor volume = length × width × 0.5 mm3). The fluorescence of pulmonary metastasis tumor was measured by FUSION FX7 Spectra Imaging system (Vilber, France).
5
Evaluating BMP-9 Treatment in Mice
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Male BALB/c-nu mice (16–18 g) were obtained from the Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). All experimental procedures and protocols were approved by the Experimental Animal Ethics Committee of Academy of Military Medical Science of China (approval no. IACUC-2017246). Mice were randomly divided into two groups (n = 7), and AdBMP-9 or AdGFP cells were injected subcutaneously into the dorsal fank of the nude mice. The tumor volume and bodyweight were measured on alternate days. All mice were sacrificed on the 30th day.
6
BALB/c-nu Mice Breeding Protocol
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Male BALB/c-nu mice were purchased from Vital River, Peking, China, and were bred in-house under specific pathogen-free (SPF) conditions. The animal experiments were performed according to the guidelines of the Animal Care and Use Committee of The Third Affiliated Hospital of Sun Yat-sen Universityand approved by the Animal Care and Use Committee of The Third Affiliated Hospital of Sun Yat-sen University.
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