Female nude mice (5-week-old, 17.9 ± 0.82 g) were purchased from Shanghai Laboratory Animal Center (SLAC, Shanghai, China). All animal experimental procedures were performed in accordance with the guidelines defined by the ethics committee and ethics committee approval. The RT4 cells were transfected with siRNA targeting LINC00467, overexpression plasmids, or vector plasmids. Stably transfected bladder cancer cells were collected during logarithmic growth phase. Then, the bladder cancer cells were washed and resuspended in PBS to achieve a cell concentration of 1 × 107 cells/ml. A 200 μL cell suspension was subcutaneously implanted into the right armpit of nude mice (the nude mice were divided randomly into three groups of 12), and macroscopic tumors were grown in the mice. The size of the tumor was calculated by measuring its length (L) and width (W) every 3 days. The nude mice were sacrificed after 25 days, and tumor volumes were measured according to the formula V = 1/2 (L × W2). All experiments were approved by the Ethics Committee of the Third Xiangya Hospital of Central South University.
Female nude mice
Manufactured by Shanghai Laboratory Animal Center
Sourced in China
Female nude mice are laboratory animals that lack a functional immune system due to a genetic mutation. They are commonly used in biomedical research to study various diseases and test new treatments, as their lack of immune response allows for the study of human diseases and therapies without interference from the animal's immune system.
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7 protocols using female nude mice
1
Bladder Cancer Xenograft in Nude Mice
2
Xenograft Model of A549 Lung Cancer
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Female nude mice were obtained from Shanghai Experimental Animal Center and maintained in pathogen-free conditions. Exponential phase A549 cells were trypsinized and washed with PBS (HyClone) and suspended in fresh PBS in 4°C. 5X106 cells were injected subcutaneously into four-week-old Female nude mice on both left and right flank. Tumor size was measured every 7 days, and tumor volume was estimated using the formula: tumor volume=0.5×length×width2. At the end of this experiment, tumors were harvested, weighed and photographed. All procedures for animal experimentats were performed in accordance with the Institutional Animal Care and Use Committee guidelines of the Animal Core Facility of the Institutes of Biochemistry and Cell Biology (SIBCB). The approval ID for using the animals was 087 by the Animal Core Facility of SIBCB.
All the experiments were repeated more than twice.
All the experiments were repeated more than twice.
3
Standardized Mouse Housing Conditions
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Female C57BL/6 mice and female nude mice (6–8 weeks old) were obtained from the Shanghai Laboratory Animal Center (Shanghai, China). Briefly, mice were fed with free access to pellet food and water in plastic cages at 21±2 °C and kept on a 12-h light–dark cycle. Animal welfare and experimental procedures were carried out strictly in accordance with the Guide for the Care and Use of Laboratory Animals (The Ministry of Science and Technology of China, 2006) and the related ethical regulations of our university. All efforts were made to minimize animal suffering and to reduce the number of animals used.
4
Investigating DGCR5 knockdown in nude mouse xenograft model
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Female nude mice (4 weeks, 18-22g) were purchased from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences (Shanghai, China). Mice were randomly divided into 2 groups and housed in appropriate environment with abundant food and water. NOZ cells infected with LV-NC or LV-shDGCR5 were selected by applying puromycin to construct stable transfected cells. Knockdown efficiency of DGCR5 was estimated by GFP expression intensity and further tested by qRT-PCR. After a week of adjustable feeding, NOZ cells (2 × 106 in 100 μL PBS) transfected with LV-NC or LV-shDGCR5 were subcutaneously injected into the left axilla of the mouse. The tumor volumes were estimated weekly (0.5 × width2 × length) by caliper. After 4 weeks, mice were sacrificed by dislocation and the tumors were collected and weighed. Total RNA and proteins were extracted from the 2 tumor groups and the rest of the tumors were stored in 4% paraformaldehyde for further assays. This animal study was approved by the Ethics Committee of Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine.
5
Xenograft Tumor Model in Mice
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Female nude mice (4 weeks old) and female BALB/c mice (5–6 weeks) were purchased from Shanghai Laboratory Animal Center and housed in a specific pathogen-free environment according to the Ethics Committee for Animal Studies of Zhejiang University (SRRSH202107012, Hangzhou, China). Stable transfected MDA-MB-231 cells (1×10*6 per mouse) or 4T1 cells (0.5×10*6 per mouse) were injected into the right subaxillary region.
Mice were monitored and measured twice per week using a slide caliper including tumor length (L), and width (W). Tumor volume (mm3) = π/6×length × width.2 (link) Mice were then sacrificed and some tumor tissues were fixed with 10% paraformaldehyde for immunohistochemical analysis and western analysis. Animal care and experiments involved in this study were performed in accordance with Accreditation of Laboratory Animal Care International guidelines.
Mice were monitored and measured twice per week using a slide caliper including tumor length (L), and width (W). Tumor volume (mm3) = π/6×length × width.2 (link) Mice were then sacrificed and some tumor tissues were fixed with 10% paraformaldehyde for immunohistochemical analysis and western analysis. Animal care and experiments involved in this study were performed in accordance with Accreditation of Laboratory Animal Care International guidelines.
6
Celastrol and 19-048 Inhibit Tumor Growth in Nude Mice
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Female nude mice (4-6 weeks old) were purchased from the Shanghai Laboratory Animal Center, Chinese Academy of Sciences. SW620 cells (2 × 106/mice) were subcutaneously injected into mice. When most of the tumors reached approximately 50 mm3, the mice were divided into 3 groups based on the tumor volume, and each group of mice were intraperitoneally injected with vehicle (2% DMSO, 5% PEG-400, 5% Tween-80), 2 mg/kg Celastrol or 2 mg/kg 19-048 5 days per week for 2 weeks (totally 10 times). The tumor size was measured using a caliper three times a week, and the tumor volume was calculated with the formula V = 1/2 (length × width2). After two weeks, the mice were sacrificed and the tumors were collected and photographed. Tumor tissues were embedded in paraffin wax, followed by immunohistochemistry. Animal experiments were handled with all of the relevant ethical regulations related to animal research. The study was approved by the Institutional Animal Care and Use Committee at Shanghai Jiao Tong University School of Medicine.
7
Extracellular Vesicle-Mediated Tumor Suppression
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Female nude mice (5-6 weeks old) were purchased from the Shanghai Experimental Animal Center and raised under aseptic laminar flow conditions. The mice were reared at 50-60% humidity and 25°C, and maintained in a 12 h light/dark cycle. Food and water were provided a Extracellular vesicles d libitum.
Each mouse was injected with 2×107 LY8 cells into the right abdomen. When a measurable tumor (approximately 100 mm3) was observed at the injection site after 3 weeks, the mice were subjected to drug treatment. Based on body weight, the mice were randomly allocated into 3 groups, with 10 mice in each group: The LY8 group (injected with LY8 cells + 10 mg/kg rituximab), GW group (injected with the same dose of GW4869 medium + 10 mg/kg rituximab), and EV group (injected with 30 µg EVs + 10 mg/kg rituximab). Five mice in each group were injected once a week and their tumors were measured every 4 days to record the long (A) and short (B) diameters of the tumors. The maximum diameter and volume of the tumor observed in the mice were 2 cm and 3.4 cm2, respectively. On the 28th day, the mice were sacrificed through cervical dislocation and tumors were removed. The precise size of the tumor was measured using a Vernier caliper, and the tumor was weighed.
Each mouse was injected with 2×107 LY8 cells into the right abdomen. When a measurable tumor (approximately 100 mm3) was observed at the injection site after 3 weeks, the mice were subjected to drug treatment. Based on body weight, the mice were randomly allocated into 3 groups, with 10 mice in each group: The LY8 group (injected with LY8 cells + 10 mg/kg rituximab), GW group (injected with the same dose of GW4869 medium + 10 mg/kg rituximab), and EV group (injected with 30 µg EVs + 10 mg/kg rituximab). Five mice in each group were injected once a week and their tumors were measured every 4 days to record the long (A) and short (B) diameters of the tumors. The maximum diameter and volume of the tumor observed in the mice were 2 cm and 3.4 cm2, respectively. On the 28th day, the mice were sacrificed through cervical dislocation and tumors were removed. The precise size of the tumor was measured using a Vernier caliper, and the tumor was weighed.
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