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Tumorigenicity Assay in Nude Mice
Published: Vol 7, Iss 13, Jul 5, 2017 DOI: 10.21769/BioProtoc.2364 Views: 21833
Edited by: Antoine de Morree
Original research article
The authors used this protocol in:
Aug 2015
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Abstract
Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and the tumor growth is monitored over a period of observation or treatment. When tumor grows to a pre-determined size or by the end of the limited period, the nude mice will be euthanatized and the xenograft will be removed for further examination.
Keywords: Tumorigenicity
Nude mice
Tumor cell transplantation
Tumor xenograft model
Background
With high incidence and mortality, tumor is one of the leading causes of death and is a major public health problem. Extensive studies are conducted every year to explore tumor pathogenesis and anti-tumor therapy. In the process of cancer research, tumorigenicity assay in nude mice is a widely used experiment to monitor tumor growth in vivo (Giovanella et al., 1974). The most commonly used animal system for tumorigenicity assay is athymic nude mouse (Petricciani et al., 1973) where malignant cells can be transplanted either subcutaneously or subrenal-capsularly (van Meir, 1997). For cells with relatively low ability of tumorigenicity, take rate can be improved by irradiation (30-60 Gy) or implantation in Matrigel (Pretlow et al., 1991). Tumorigenicity assay provides a means of generating human cell derived tumor tissues for measurement of tumor cell malignancy and evaluation for anti-tumor drug efficacy.
Materials and Reagents
- 0.1-20 ml pipette tips (Eppendorf, catalog number: 22492012 )
- 5-200 ml pipette tips (Eppendorf, catalog number: 22492039 )
- 50-1,000 ml pipette tips (Eppendorf, catalog number: 22492055 )
- Cell culture disc (75-cm2) (Corning, catalog number: 430641 )
- Falcon 15 ml conical centrifuge tubes (Corning, catalog number: 430791 )
- Counting slides (Bio-Rad Laboratories, catalog number: 1450011 )
- Eppendorf tubes (1.5 ml) (Eppendorf, catalog number: 0030120086 )
- Tuberculin syringe (1 ml) (BD, catalog number: 300841 )
- BD 1 ml syringe (BD, catalog number: 309628 )
- BD PrecisionGlide needle (25 G, 0.5 x 40 mm) (BD, catalog number: 301808 )
- Female athymic nude mice (6-8 week old, Fourth Military Medical University, Experimental Animal Center, BALB/c)
- Phosphate buffered saline (PBS) pH 7.4 (Thermo Fisher Scientific, GibcoTM, catalog number: C10010500BT )
- Trypsin-EDTA (0.25%) (Thermo Fisher Scientific, GibcoTM, catalog number: 25200072 )
- Ethanol (Tianjin Fuyu Fine Chemical, catalog number: 20160916 )
- RPMI 1640 medium (Thermo Fisher Scientific, GibcoTM, catalog number: C11875500BT )
- Fetal bovine serum (FBS) (Thermo Fisher Scientific, GibcoTM, catalog number: 10099141 )
- Penicillin-streptomycin (5,000 U/ml) (Thermo Fisher Scientific, GibcoTM, catalog number: 15070063 )
- L-glutamine (Thermo Fisher Scientific, GibcoTM, catalog number: 25030081 )
- Complete 1640 medium (see Recipes)
Equipment
- 2-20 μl pipettes (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 4641060N )
- 20-200 μl (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 4641080N )
- 100-1,000 μl (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 4641100N )
- Water-Jacketed CO2 incubators (Thermo Fisher Scientific, model: Model 3100 Series , catalog number: 3131)
- Clean bench (Thermo Fisher Scientific, model: HeraguardTM ECO )
- Centrifuge (Eppendorf, model: 5424 R )
- Automated cell counter (Bio-Rad Laboratories, model: T20TM )
- Microscope (Olympus, models: CX23 and BX53 )
- Caliper (Mitutoyo, catalog number: 530-118 )
Procedure
Category
Cancer Biology > Cancer stem cell > Animal models > Cell invasion
Cell Biology > Cell Transplantation > Allogenic Transplantation
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