Four-week-old male nude mice were bought from GemPharmatech company and raised in a pathogen-free barrier environment. HCC cells with or without CAFs were resuspended in PBS and slowly injected into the left lobe of the liver. Animals were sacrificed about 4–5 weeks after implantation. Bioluminescence was measured after an intraperitoneal injection of 100 µl of potassium d -luciferin salt (30 mg/ml, per animal). All experiments were approved by Tongji Hospital Institutional Review Board (IRB ID: TJH-202206023).
Nude mice
Manufactured by GemPharmatech
Sourced in China
Nude mice are a strain of laboratory mice that are genetically modified to lack a functional immune system. They are commonly used in biomedical research and drug development for the study of human diseases, as their lack of an adaptive immune response allows for the transplantation and growth of human cells and tissues.
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8 protocols using nude mice
1
Liver Tumor Xenograft Modeling in Mice
2
Nude Mice Tumor Xenograft Model
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Twenty 4-week-old female nude mice were purchased from Gempharmatech Co., Ltd and were randomly divided into four experimental groups. They are kept at the Animal Research Centre of Tongji Medical College, Huazhong University of Science and Technology. We inoculated SiHa cells overexpressing or knock-down HCP5 (SiHa-lv-hcp5, SiHa-lv-shhcp5) into the right hindquarters of nude mice subcutaneously to observe tumorigenesis, and the control group was inoculated in the same way. All the nude mice were killed, and tumor tissue was collected after 30 days for comparative analysis. The length and width of the tumor were measured, and the volume is calculated using the formula as follows: [(length x width)2]/2. Tumor weight was also measured. Our animal experiment research was carried out strictly in accordance with the “Guidelines for the Welfare of Experimental Tumor Animals”. All experimental protocols were approved by the Animal Care and Use Committee of Huazhong University of Science and Technology.
3
Nude Mice Xenograft Tumor Study
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6- to 8-week-old female nude mice were purchased from GemPharmatech, Nanjing, China. Neuro2a xenograft was established via axillary inoculation in nude mice. When tumors reached an average volume of 30–45 mm3, mice were randomized into control and treated groups. Mice were orally administered (i.g.) with placebo (CMC-Na), 40 and 80 mg/kg of Hu7691, and 80 mg/kg of ATRA, respectively. The dosage and dosing frequency of each group was based on the data from preclinical trials. Tumor volume and body weight were measured every day. On the 17th day of treatment, mice were sacrificed, and tumor samples were divided into two pieces and kept at −80 °C for protein and RNA extraction.
4
Targeting circRNA-0007919 in GEM-Resistant Pancreatic Cancer
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PANC-1 and CFPAC-1 GEM-resistant cells were infected by hsa_circ_0007919 inhibition lentivirus (GenePharma, China) or negative control lentivirus and selected by puromycin (Solarbio, China) for over 2 weeks, the efficiency of lentivirus was detected by qPCR. 5 × 106 lentivirus-infected cells were injected into blank region of nude mice (Gempharmatech, China) and were treated with GEM (50 mg/kg, i.p.) every 4 days. After measuring volumes of tumors every 5 days, the mice were sacrificed and the tumors were harvested 25 days after injection. All sequences of shRNAs are shown as follows:
sh-hsa_circ_0007919: 5’-CACCGAGGTGGAAGCAGGGAAAGTTCGA AAAAATTGATCAATGCCGAGGA-3’;
sh-Ctrl: 5’-CACCGTTCTCCGAACGTGTCACGTTTCGAAAAACGTG ACACGTTCGGAGAA-3’.
sh-hsa_circ_0007919: 5’-CACCGAGGTGGAAGCAGGGAAAGTTCGA AAAAATTGATCAATGCCGAGGA-3’;
sh-Ctrl: 5’-CACCGTTCTCCGAACGTGTCACGTTTCGAAAAACGTG ACACGTTCGGAGAA-3’.
5
Subcutaneous Mouse Tumor Modeling
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Five to 6-week-old female C57bl/6 mice, male BALB/c, and nude mice were purchased from GemPharmatech (Nanjing, China). Mice were maintained at a specific pathogen-free facility with a 12 h/12 h day/night turnover. All animal experiments were approved by the Research Ethical Committee of the Sixth Affiliated Hospital of Sun Yet-sen University and in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory animals. For the subcutaneous mouse model, 2 × 105 MC38 or 1 × 105 CT26 with IFI35 stable knockdown or IFI35 overexpression constructs, or control cells suspended in 0.1 mL PBS were injected into the flanks of the mice. Tumor growth was assessed by caliper measurement three times in 1 week. The following formula was used to calculate tumor volume: Tumor volume (mm3) = (tumor width2 × length)/2. On day 7 after tumor cell injection, anti-PD-1 (InVivoMab) or IgG1 isotype monoclonal antibodies (InVivoMab) were intraperitoneally injected at a dose of 200 µg per mouse every 3 days for the duration of the experiment.
6
Xenograft Tumor Model in Nude Mice
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Four- to five- week-male nude mice were purchased from Gem Pharmatech Co., Ltd and housed on a 12 h light–dark cycle under pathogen-free conditions. The animal experiment was approved by the Committee on the Ethics of Animal Experiments of the Yantai Institute of Materia Medica, and was performed in strict accordance with the guidelines of the National Research Council's Guide for the Care and Use of Laboratory Animals. HCT-116 cells were harvested, 2 × 106 (link) cells per 200 μL were injected on the flank of the mice. Tumor growth was measured, when the average tumor volume reached to 200 mm3, the mice were randomized into six groups (n = 5). The mice in the treatment groups were treated with DM1 (1 mg kg−1), LWJ-M30 (1 mg DM1 equiv. kg−1) twice a week in the tail. The mice in control group were administrated with 5% glucose solution. Tumor size was measured three times a week using a caliper and the volume was calculated as
After 21 days treatment, the mice were euthanized and the tumors were excised and weighed.
After 21 days treatment, the mice were euthanized and the tumors were excised and weighed.
7
Subcutaneous Tumor Xenograft Assay
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The indicated cells were subcutaneously injected into the dorsal flank of 4-week-old nude mice (GemPharmatech, Nanjing, China). After 21 days, mice were sacrificed and tumors were excised and weighed. Studies on animals were conducted with approval from the Animal Research Ethics Committee of Anhui Medical University (LLSC20200763).
8
Xenograft Model for Evaluating NEAT1 Silencing
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Nude mice were purchased from GemPharmatech (Chengdu, China). All animal experiments were performed according to Chongqing University Animal Care Guidelines. For subcutaneous xenografts, HepG2 cells stably silencing NEAT1 (5 × 106) were injected into 6-week-old female Nude mice. Tumor length (L) and width (W) were monitored weekly. Tumor volume was calculated through (L × W2)/2 [39 (link)]. Tumors were then harvested and fixed in 4% PFA.
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