Athymic BALB/c mice (4-6 weeks old) were obtained from Charles River (Beijing, China) and maintained under pathogen-free conditions. All mice were raised at a 12 h dark/light cycle, at 24 ± 2 °C, and were given free access to food and water. Animal experiments were carried out in approval of the Experimental Animal Committee of Panyu District Central Hospital (PYRC-2021-113), and conducted under the 3R principle to reduce suffering of mice. SW480 cells (5 × 106) treated with miR-491-3p mimic, control miRNA mimic or both miR-491-3p mimic and pcDNA-uMtCK were resuspended in 100 µL of PBS and injected into right flank of mice (n = 3). Tumor growth was half-quantified with a caliper in a blinded fashion at the indicated time (0, 14, 21, and 35 days). Tumor volume was estimated through the formula length ×width2 ×0.5, as previously described (Hart et al., 2012 (link); Khare et al., 2019 (link)). At the end of studies, all mice were euthanatized by cervical dislocation under inhalation anesthesia (2% isoflurane), and tumor tissues were collected. During the experiment, once any mouse suffering from unexpected disease or over-sized tumor (>1 cm) will be euthanatized as described earlier.
Balb c athymic mice
Manufactured by Charles River Laboratories
Sourced in China
BALB/c athymic mice are an inbred strain of laboratory mice characterized by a congenital absence of a functional thymus gland, resulting in a deficiency of T cells. This mouse model is widely used in biomedical research, particularly in the fields of immunology and oncology.
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15 protocols using balb c athymic mice
1
In Vivo Evaluation of miR-491-3p and uMtCK in Tumor Growth
2
Assessing Transformation Potential of MEFs
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For focus assay, MEFs were seeded at 150, 300 or 600 cells/well in six-well plates. After 2 weeks, cells were stained with crystal violet and colonies more than 2mm in diameter were counted under a Nikon-Eclipse TS100 light microscope. The experiments were repeated twice with two independent MEFs preparations. For soft agar assay, cells were resuspended in 0.3% agarose in DMEM at a density of 5×104 cells/well in six-well plates and plated in triplicate onto solidified 0.6% agarose-containing medium. Cells grew for 4 weeks and were supplemented with fresh media two times a week. Colonies were stained by 0.005% crystal violet and counted using a light microscope. The experiments were repeated twice with different MEFs preparations. For in vivo tumorigenesis assays, 5×106 cells were implanted s.c. into each flank of five 8-week-old female athymic BALB/c mice (Charles River Laboratories). Tumor growth was monitored with a caliper for 3 weeks. No tumor developed in control group during the period of observation of 10 weeks. The experiment was repeated once.
3
Overexpression of MTA2 Drives Tumor Growth
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KYSE30 cells were used in this study to establish stable cell lines via virus transfection that constitutively overexpresses the MTA2 protein or MTA2 shRNA. Four‐wk‐old male athymic BALB/c mice (Charles River) were randomly divided into 2 groups. For the tumor formation assay, 1 × 106 KYSE30/shNC or KYSE30/shMTA2 and KYSE30/CD511B or KYSE30/MTA2 cells in 100 μL PBS were injected subcutaneously into the right flank of each mouse. Tumor nodules were measured every 7 d after their length exceeded 4 mm, and the volume was calculated using the following formula: V = (width2 × length)/2. Xenografts were collected at the 6th wk for immunohistochemical staining and protein extraction.
For pulmonary metastasis models, 5 × 106 KYSE30/shNC or KYSE30/shMTA2 cells in 200 μL PBS were injected via the tail vein. All mice were sacrificed 2 mo later. Pulmonary metastases were examined in the gross specimens and with H&E staining of lung tissues.
For pulmonary metastasis models, 5 × 106 KYSE30/shNC or KYSE30/shMTA2 cells in 200 μL PBS were injected via the tail vein. All mice were sacrificed 2 mo later. Pulmonary metastases were examined in the gross specimens and with H&E staining of lung tissues.
4
Xenograft tumor growth in athymic mice
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Athymic BALB/C mice (4-6 wk old) were purchased from the Beijing Vital River Laboratory Animal Technology Co., Ltd (Beijing, China) and maintained in a SPF facility. Huh7 cells (5 × 106) over-expressing lncRNA W5 were subcutaneously injected into the flanks of nude mice. Tumor length (L) and width (W) were measured using calipers every 3 d up to 6 wk. Tumor volume was estimated using the formula: π × length × width 2/6. After 6 wk, the mice were sacrificed, and tumor volumes and weights were examined. Proliferation progression was examined and quantified using a noninvasive bioluminescence in vivo Imaging System (Xenogen Corporation, Alameda, CA, United States). All animal experiments were conducted with the approval of the Fifth Medical Center of Chinese PLA General Hospital’s Animal Care and Use Committee.
5
Xenograft Tumor Growth in Athymic Mice
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Four-week-old female athymic BALB/c mice were purchased from Vital River Laboratories. For xenograft models, 2 ´10 6 HeLa cells transfected with siNC or siHOTAIR were subcutaneously injected in the right ank of BALB/c nude mice (n=5 per group). Three weeks later, mice were sacri ced, tumors were homogenized, and proteins were extracted for western blotting. All animal procedures were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee at the Institute of Wuhan University of Science and Technology.
6
Xenograft Tumor Growth Assay for LINC01224
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sh-LINC01224#1 or sh-NC-transfected SW620 cells (5 × 106) were subcutaneously injected in 5-week-old male BALB/c athymic mice (Vital River Laboratory Animal Technology, Beijing, China) (n = 6/group). The length and width of each xenograft tumor were monitored every 7 days, and tumor volume was calculated by 0.5 × length × width2 [31 (link)]. After 35 days, tumor-bearing mice were euthanized using 5% isoflurane, and the tumors were detached and weighted. And, tumor tissues were lysed to detect relative levels of LINC01224, miR-485-5p, and MYO6.
7
BALB/c Athymic Mouse Experiment
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All animal experiments followed the protocol approved by the Animal Research Management Committee of Shandong University. All methods are reported in accordance with ARRIVE guidelines (https://arriveguidelines.org ) for the reporting of animal experiments and all methods were carried out in accordance with the relevant guidelines and regulations. Male BALB/c athymic mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All animal procedures followed the instructions of the Institutional Animal Care and Use Committee (IACUC) of Shandong University (Jinan, China).
8
Targeting SOX2 in Ewing's Sarcoma Xenografts
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BALB/c athymic mice at 6–8 weeks old were acquired from Vital River (Beijing, China) and kept according to animal welfare guidelines of Peking University People’s Hospital. Each mouse was injected in the right flank with 3 × 106 A673 Ewing’s sarcoma cells. When the xenograft tumors were palpable, the mice were randomly divided into three groups. For each group, intratumoral injection with siSOX2, siNC or glucose was performed once every 2 days. Five microgram siSOX2 or siNC was mixed with 8 μl transfection reagent (Entranster-in vivo; Engreen, Beijing, China) and hydrated with 10 % glucose injection at a concentration of 50 μg/ml for each injection. The tumor volumes were calculated ([length × width2]/2) every 3 days and the data were plotted for comparison. After 30 days, the mice executed humanely and the tumors were excised and photographed. Xenograft tissue samples were prepared for subsequent IHC and Western blot assays.
9
Monitoring Breast Cancer Metastasis in Mice
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MCF7 Tet-Off p95HER2 cells expressing a short hairpin control or targeting ADAM17 were co-injected with MDA-MB-231 Luc cells into the right flanks of 6- to 8-week-old female BALB/c athymic mice (Charles River Laboratories, Paris, France) in addition to a 17β-estradiol pellet (Innovative Research of America, Sarasota, FL, USA). The expression of p95HER2 was repressed by adding doxycycline (50 mg/kg per day) to the drinking water until tumors were about 100 mm3. Tumor xenografts were measured with calipers every 3 days, and tumor volume was determined by using the formula: (length × width2) × (pi/6). Tumors were resected when they reached 300 mm3, and metastatic colonization was monitored by in vivo bioluminescence imaging with the IVIS-200 imaging system (PerkinElmer, Waltham, MA, USA). At the end of the experiment, animals were anesthetized with a 1.5 % isofluorane-air mixture and were sacrificed by cervical dislocation. Mice were maintained and treated in accordance with institutional guidelines of Vall d’Hebron University Hospital Care and Use Committee.
10
Curcumin Inhibits Tumor Growth in Xenograft Model
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Total of 20 BALB/c athymic mice (female, 5-week-old, ~ 20 g) were purchased from Charles River Laboratories (Beijing, China), and housed in specific pathogen-free microisolator cages. A2780 cells (3 × 106) transfected with oe-circ-PLEKHM3 or lentiviral vector were injected into mice (n = 5/group) via subcutaneous inoculation. After 1 week, mice were intraperitoneally injected with 15 mg/kg of curcumin or equal volume of DMSO every 2 days. Tumor volume was monitored every week, and analyzed according to the formula volume = 0.5 × length × width2. After 5 weeks, mice were euthanized using 5% isoflurane. Tumor tissues were collected and weighed. The animal experiments were in line with the guidelines of the National Institutes of Health guide for the Care and Use of Laboratory animals (NIH Publications No. 8023, revised 1978), and approved via the Institutional Animal Care and Use Committee of The Second Hospital of Dalian Medical University.
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