To determine the in vivo tumorigenicity, subcutaneous animal models were established. A total of 18 male BALB/c athymic nude mice, aged 4–6 weeks, body weight 15–20 g were purchased from Vital River Laboratories Co., Ltd. (Beijing, China), provided with sterilized water and food, and housed in standard lab conditions with 12 h light/dark cycles. The procedures for the care and use of animals followed the guide for the Care and Use of Laboratory Animals (24 ) and experimental protocols were approved by the Animal Care and Use Ethics Committee of Shenzhen University. The mice were randomly divided into three groups (n=6 per group) according to the injected cells. Following lentivirus infection and selection, the U2OS cells found to stably express miR-124 (miR-124) and empty vector cells (miR-NC) were washed and re-suspended with PBS; 1×106 cells (200 µl) were then subcutaneously injected into the dorsal flank of the nude mice. The mice were monitored daily for 1 month following injection, and the size of the xenografted OS tissues and the tumor formation rate of the three groups were compared. After 22 days, the mice were sacrificed and the tumors were dissected and weighed.
Balb c athymic nude mice
Manufactured by Charles River Laboratories
Sourced in China, Japan
BALB/c athymic nude mice are a strain of laboratory mice that lack a functional thymus gland. This results in a deficiency of T cells, leading to a compromised immune system. These mice are commonly used in research applications that require an immunodeficient animal model.
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Lab products found in correlation
Matrigel, by BD (3 mentions)
C57bl 6 mice, by Charles River Laboratories (2 mentions)
Balb c mice, by Charles River Laboratories (2 mentions)
Scid mice, by Jackson ImmunoResearch (2 mentions)
Nsg mice, by Jackson ImmunoResearch (2 mentions)
Lv3 cmv gfp puro vector, by GenePharma (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Female balb c mice, by Charles River Laboratories (1 mentions)
Sodium acetate anhydrous ch3coona, by Merck Group (1 mentions)
Mir 100 5p agomir, by RiboBio (1 mentions)
Ivis lumina xrms series 3 imaging system, by PerkinElmer (1 mentions)
Matrigel basement membrane matrix, by BD (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Formalin, by Merck Group (1 mentions)
Balb c nude mice, by Charles River Laboratories (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
Matrigel, by Corning (1 mentions)
17β estradiol pellet, by Innovative Research (1 mentions)
Accell sirna, by Horizon Discovery (1 mentions)
60 protocols using balb c athymic nude mice
1
In Vivo Tumorigenicity Assessment of miR-124 in Osteosarcoma
2
Xenograft Assays for Prostate Cancer
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Xenograft assays were performed as we previously reported 26 (link). We used 20 four-week-old male BALB/c athymic nude mice (Vital River, Beijing, China) to establish xenograft models. N-cadherin-overexpressing PC3 (PC3-sh-CDH2) cells or negative control (PC3-sh-NC) cells (1 × 107) were mixed with 200 μl of phosphate-buffered saline (PBS) containing 30% Matrigel (BD Biosciences). The cells were subcutaneously inoculated into the right axilla of mice, and the mice were intraperitoneally injected with 25 mg/kg/day enzalutamide (ENZ) or the same volume of dimethyl sulfoxide (DMSO). Tumor size was measured every 5 days with a caliper, and tumor volume was calculated using the following formula: volume (mm3) = (length × width2) × 0.5. Fifty days post-inoculation, the mice were euthanized, and pathological analyses of the dissected tumors were performed. All experiments were performed in accordance with the institutional ethical guidelines of Capital Medical University.
3
Lentivirus-Mediated Tumor Progression in Nude Mice
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Lentivirus encoding TAF15 and LINC00665 was generated with a pLenti6.3/V5eDEST Gateway vector kit (Life Technologies). sh-MTF1 and sh-YY2 were ligated into the LV3-CMV-GFP-Puro vector (GenePharma), respectively. Thus, target lentivirus vectors were generated to obtain the stable expressing cells of precursor (pre-)TAF15, pre-LINC00665, and sh-MTF1 or sh-YY2. Four-week-old BALB/c athymic nude mice were purchased from Beijing Vital River Laboratory Animal Technology (Beijing, China). These nude mice were divided into seven groups: control, TAF15+, LINC00665+, MTF1−, YY2−, TAF15+ + LINC00665+ + MTF1−, and TAF15+ + LINC00665+ + YY2− groups. 3 × 105 (100 μL) cells were injected subcutaneously into the right limb of nude mice. We observed and recorded the tumor formation time and tumor formation of nude mice in each group, calculating the tumor formation rate and detecting the weight and volume of transplanted tumor to draw the tumor growth curve [tumor volume (mm3) = (longest diameter × shortest diameter)2 × 0.5]. The number of survived nude mice was recorded, and survival analysis was performed using a Kaplan-Meier survival curve.
4
Angiogenesis Assay in Nude Mice
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Four-week-old male BALB/C athymic nude mice were purchased from the Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). The animals were fed with autoclaved food and water during the experiment. All animal procedures were performed in strict accordance with the protocol approved by the Administrative Panel on Laboratory Animal Care of China Medical University (Shenyang, China). In brief, 3 × 106 GECs resuspended in 400 μL solution containing 80% Matrigel were subcutaneously injected. Plugs were harvested after 4 days and then weighed, photographed, and dispersed in 400 μL PBS (with overnight incubation at 4 °C) to collect the hemoglobin. Hemoglobin content was measured using Drabkin’s reagent solution (Sigma-Aldrich) according to the manufacturer’s instructions.
5
Xenograft Model for PSCA Knockdown in Gastric Cancer
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The Research Animal Resource Center of Sun Yat-sen University approved the use of laboratory animals in this study. BALB/c athymic nude mice (4 weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). The experiments and procedures on animals conformed to the institutional animal care guidelines. A xenograft model was used. The mice were injected with 5 x 106 MKN45 cells or MKN45-shPSCA cells in the right subcutaneous armpit. Tumour size was measured twice a week. Mice were sacrificed after the size of the tumour exceeded 1000 mm3.
6
Hydrogen therapy for lung cancer
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Animal experiment was performed in the Third Hospital of Hebei Medical University by Wang et al. [8 (link)] in accordance with National Institute of Health’s Guidelines for the Care and Use of Laboratory Animals, and was given permission by Animal Care and Research Committee of the Third Hospital of Hebei Medical University. Four-week male BALB/c athymic nude mice (Beijing Vital River Laboratory Animal Technology, Beijing, China) were fed in specific pathogen-free conditions. A total of 1 × 107 A549 cells suspended in 200 μl PBS were injected into the BALB/c mice, which were then divided into three groups when the tumors were grown 3–4 mm in diameter, those were H2 group, cis-platinum group and control group. Mice in H2 group were given 60% H2 inhaling for 2 h every day, mice in cis-platinum group were given cis-platinum (10 mg/kg body weight) [17 (link)] intraperitoneal injection. H2 and cis-platinum treatments were kept for 4 weeks. Then, the mice were killed via cervical dislocation. Atmosphere was used as a control of 60% H2 and the same amount of saline was served as a control for cis-platinum.
7
Synthesis and Evaluation of Theranostic Nanoparticles
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All chemicals of analytical grade were used without further purification. Chloroauric acid (HAuCl4·4H2O) was purchased from Chemart (Tianjin, China). Reduced glutathione (rGSH, 98%) was obtained from Solarbio (Beijing, China). Diethylenetriamine-pentaacetic acid dianhydride (DTPAA, 95%) was purchased from Alfa Aesar (Shanghai, China). Gadolinium (Ⅲ) chloride (GdCl3·6H2O), Dimethyl sulfoxide (DMSO, >99%), and Sodium acetate anhydrous (CH3COONa) were purchased from Sigma-Aldrich (St. Louis, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Merck (Germany). The other chemicals were purchased from Sigma-Aldrich (St. Louis, USA). Deionized water used throughout the experiments was acquired from Mili-Q water purification system after its resistivity reached 18.2 MΩ CM.
Human renal epithelial cell line (293T cell line) and human hepatocellular carcinoma cell line (HepG2 cell line) were acquired from Molecular Imaging Lab of Tianjin Medical University (Tianjin, China). The use of these cell lines had Molecular Imaging Lab of Tianjin Medical University review board approval. The female BALB/c mice and BALB/c athymic nude mice were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd (Beijing, China).
Human renal epithelial cell line (293T cell line) and human hepatocellular carcinoma cell line (HepG2 cell line) were acquired from Molecular Imaging Lab of Tianjin Medical University (Tianjin, China). The use of these cell lines had Molecular Imaging Lab of Tianjin Medical University review board approval. The female BALB/c mice and BALB/c athymic nude mice were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd (Beijing, China).
8
Xenograft Assay of OIP5 Knockdown
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SW780 cells stably transfected with OIP5 knockdown vector (miRNA-OIP5) and its negative control (miRNA-NC) were employed to the tumorigenicity assay. Five female BALB/c athymic nude mice (Five-six weeks old, 16-20g) were purchased from Beijing Vital River Laboratory Animal Technology Co, Ltd. All mice were housed in a climate-control SPF (Specific Pathogen Free) facility. Cell suspension (1× 107 cells in 200 μl PBS per mouse) was subcutaneously injected into the flanks of the nude mice. And, the miRNA-OIP5 cell line was injected on the right side, and the miRNA-NC cell line was injected on the left side, respectively. The tumor size, volume and mice weight were measured twice a week. The tumor volume was calculated using the formula: length × width2 × 1/2. After inoculation 4 weeks, the mice were euthanized and the tumors were removed and weighed.
9
Balb/c Athymic Nude Mice Acclimation
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Four- to five-week-old male BALB/c athymic nude mice were acquired from the Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China) and acclimated for one week. All animals were housed in a controlled atmosphere (25 ± 1 °C at 50% relative humidity) under a 12-h light/12-h dark cycle. Animals had free access to food and water at all times. Food cups were replenished with fresh diet every day. The utilized animal experimental protocols received approved from both the Institutional Animal Care and Use Committee of the Laboratory Animal Centre of Peking University Shenzhen and the Tsinghua Shenzhen Graduate School (the permit from Peking University Shenzhen Graduate School is “YW”: the permit from Tsinghua Shenzhen International Graduate School is “ethical development no. 37 (Year 2019)”.
10
Xenograft Tumor Growth in Nude Mice
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Five-week-old male BALB/c athymic nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). The mice were housed in microisolator cages under a 12/12-h light/dark cycle (lights on at 8:00 a.m.) with food and water ad libitum. HepG2 cells stably transfected with Lv-LINC00355:8 or Lv-con (1 × 107) cells were injected subcutaneously into the right flanks of nude mice. The tumour volume was measured every 4 days using the formula (length × width × width)/2. Three weeks later, the nude mice were sacrificed, and the tumours were weighed. The animal experiments were approved by the Animal Ethics Committee of the Third Affiliated Hospital of Sun Yat-Sen University.
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