Male athymic balb c nude mice

Athymic male nude Balb/C mice (Charles River, 9 weeks) were randomly divided into three experimental groups: control, DP cells, and DP cells with EpSlKCs (n = 4, each group). The silicone chamber model was set and used as previously described [22 (link)]. Cell suspensions with 5.0 × 10⁶ DP cells and 2.5 × 10⁶ EpSlKCs (experimental group), 5.0 × 10⁶ DP cells (DP cell group), or vehicle (FAD medium, control group) were injected in the chamber. The top of the chamber was cut-off after 1 week, and the chamber was fully removed after 2 weeks. Animals were euthanized 6 weeks after surgery to harvest the wound area for histological analysis.

Male athymic Balb/c nude mice (4–5 weeks old) were purchased from Vital River Laboratory Animal Technology Corp. (Beijing, China). Animal experiments were performed according to the guidelines of the Institutional Animal Care and Use Committee of Peking University (permit no. 2011-0039).
The subcutaneous liver tumor model was established in Balb/c nude mice by injecting 1 × 10⁶ HepG2-Red-fLuc or SMMC-7721-fLuc cells into the right lower flank of mice. The orthotopic liver tumor model was generated by inoculating 1 × 10⁶ HepG2-Red-fLuc cells into the liver lobe of mice through laparotomy under isoflurane anesthesia. After the surgery, the peritoneum and skin of the mice were sutured with absorbable surgical lines, and the mice were placed in a warm box until they woke up.

Male athymic BALB/c nude mice, 4–6 weeks old, were purchased from Vital River Laboratory Animal Technology Co. (Beijing, China). The animal care and experimental protocols were approved by the institutional guidelines of Jiangsu Province and by the Animal Care and Use Committee of Nanjing Medical University. 1 × 10⁷ PC cells were resuspended in 200 μL PBS medium and were subcutaneously injected into the flank of each nude mouse. The tumors were measured weekly and the tumor volume was calculated following the formula length × width²/2. The mice were killed at 6 weeks after inoculation.

Male athymic BALB/c nude mice (aged 3–4 wk) were purchased from Vital River. Nude mice were housed in a standard environment (temperature, 23 ± 1°C; humidity, 40–70%) with a 12:12-h light/dark cycle. The experimental protocol was approved by the Institutional Animal Care and Use Committee of Ningbo University. All the animal experiments were conducted in accordance with the guideline for the Care and Use of Laboratory Animals published by the US National Institutes of Health.
To assess the effects of METH on liver cancer in vivo, 6 × 10⁶ HepG2 cells were injected subcutaneously into the right flank of nude mice. Then, mice were randomly assigned into different groups (three to five mice each) with various dosages of METH intraperitoneally injected as indicated. The mice were euthanized on days 7, 14, and 21. The xenograft tumors were isolated, photographed, and weighed. The sizes of isolated xenograft tumors were measured using a caliper. The maximum (L) and minimum (W) lengths of the tumors were measured, and the tumor size was calculated as ½LW².

Male athymic BALB/c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and maintained under specific pathogen-free conditions. All experiments were performed in accordance with institutional guidelines for animal care. 6- to 8-week-old nude mice were inoculated subcutaneously with HepG2 cells (1 × 10⁶/0.2 mL phosphate-buffered saline, n = 6–7). The length and width of the tumors were measured with calipers at the indicated time points.

Male athymic BALB/c nude mice were purchased from Vital River Laboratory Animal Technology Co. Ltd. (Beijing, China) and maintained under specific pathogen-free conditions in the National Chengdu Center for Safety Evaluation of Drugs. Mice were manipulated and housed according to protocols approved by the Institutional Animal Care and Treatment Committee of Sichuan University. A total of 1.0× 10⁶ SW480 cells were inoculated in 0.1 mL of serum-free medium subcutaneously in the right flank of 5–6-week old healthy BALB/c nude mice. When the tumors reached an average volume of about 60 mm³, the mice bearing too large or too small tumors were eliminated and the left were divided into groups for treatment. Turing the experiment, tumor size was measured using with calipers every three days, and tumor volumes were calculated according to the following formula: (Length × Width²) × 0.52.