The CRC cell lines MC38, CT26, and HT‐29 were purchased from the American Type Culture Collection (ATCC) and cultured according to the manufacturer's instructions. These cells were characterized using short tandem repeat markers by Genewiz, Inc. and were confirmed to be mycoplasma free (last tested in 2017). Note that 5‐ to 6‐week‐old BALB/c nude, C57BL/6, BALB/c, NOD/SCID, and NOD‐Prkdcem26Cd52IL2rgγem26Cd52/Gpt (NCG) mice were purchased from Gempharmatech and were housed under pathogen‐free conditions, in temperature‐controlled holding rooms (20−22°C), with 30%−34% humidity, and on a 12 h on (6 a.m.) and off light cycle (6 p.m.).
Balb c
Manufactured by GemPharmatech
Sourced in China
BALB/c is a strain of laboratory mice commonly used in biomedical research. It is an inbred mouse strain that is widely utilized for various experimental purposes.
Automatically generated - may contain errors
4 protocols using balb c
1
Characterization of CRC Cell Lines
2
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.
3
In Vivo Tumorigenicity Assay of Huh7 and MHCC-LM3 Cells
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For in vivo tumorigenicity assays, 3 × 106 cells, including Huh7and MHCC‐LM3 with the recombinant lentivirus were harvested and resuspended in 100 μl of PBS and subcutaneously injected into 4‐ to 6‐week‐old male BALB/c nude mice. Tumour volumes were measured as (tumour width2 × tumour length)/2 and recorded every 4 days. Mice were sacrificed at 36 days post‐inoculation. Finally, subcutaneous tumours were excised and further analysed. All male nude mice (BALB/c) were obtained from GemPharmatech Co. Ltd., Nanjing, China, and housed in a 12 h light/dark cycle at 23–26°C.
4
Generating Humanized NSG Mice
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All animal studies were approved by the Animal Experimental Ethical Committee of the Fudan University, Shanghai, China (No. 20200306-071). Male C57BL/6, female BALB/c, and male BALB/c-nu/nu nude mice at the age between 6- and 8-week-old were purchased from GemPharmatech, China, and maintained under a 12-h dark/12-h light cycle with food and water provided ad libitum. Humanized NSG mice were constructed by Shanghai Model Organisms, China. In brief, human CD34+ cells were isolated from cord blood and were transplanted into each 1.4 Gy-irradiated NSG mouse. After 16 weeks, the humanization of each mouse is analyzed by detecting the percentage of human CD45+ cells in all CD45+ cells in peripheral blood. Human CD45+ cell percentage greater than 25% was considered successful in modeling. All animals were randomly assigned to groups before experiments. No statistical methods were used to predetermine the sample size. The experimenter was not blind to the assignment of the groups and the evaluation of the results. No samples, animals, or data were excluded.
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