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4T1-Luc2
4T1-Luc2
規(guī)格:
貨期:
編號(hào):B161364
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 4T1-Luc2
商品貨號(hào) B161364
Organism Mus musculus, mouse
Tissue mammary
Product Format frozen 1.0 mL
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 2

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease This tumor produced mimics an animal stage IV human breast cancer.
Strain BALB/cfC3H
Applications Excellent signal/background ratio and stable Luciferase expression make this cell line ideal for in vivo bioluminescence imaging of xenograft animal model to study human cancer and monitor activity of anti-cancer drug. It also can be used in cell-based assays for cancer research.
Storage Conditions liquid nitrogen vapor phase
Tumorigenic Yes, tested in BALC/c mice
Comments This luciferase expressing cell line was derived from parental line CRL-2539 by transduction with lentiviral vector encoding firefly luciferase gene (luc2) under control of EF-1 alpha promoter. This cell line was established through single cell cloning, and the cells constitutively express high levels of enzymatically active luciferase protein, which can be detected via in vitro and in vivo bioluminescence assays. The cells should be maintained in Blasticidin (8 μg/mL) containing medium in routine cell culture. It is recommended to remove Blasticidin prior to and during the experiment procedure when the cells are injected into animals in vivo, or co-cultured with other cell types in vitro.
Complete Growth Medium The base medium for this cell line is RPMI-1640 Medium (ATCC 30-2001). To make the complete growth medium, add the following components to the base medium: 
  • Fetal bovine serum (FBS; ATCC 30-2020) to a final concentration of 10%
  • Blasticidin to a final concentration of 8 µg/mL

Subculturing
Note: The cells should not be allowed to become confluent, subculture at 80% of confluence. Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
    Cultures can be established between 2 x 104 and 4 x 104 viable cells/cm2.
  6. Incubate cultures at 37°C.
div>Interval: Maintain cultures at a cell concentration between 1 X 104 and 1.5 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Every2 to 3 days
Cryopreservation Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 1.0 x 106
Volume 1.0 mL
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Functional Tests Luciferase activity: signal to noise ≥ 1,000 RLUs
In Vitro Luminesence: 100,000 photons/cell/sec, subject to imaging and culturing conditions
Population Doubling Time approximately 12.3 hrs
Year of Origin 2018
References

Zinn KR, et al. Noninvasive bioluminescence imaging in small animals. ILARJ 49: 103-115, 2008. PubMed: 18172337

Dothager RS, et al. Advances in bioluminescence imaging of live animal models. Curr Opin Biotechnol 20: 45-53, 2009. PubMed: 19233638

Pulaski BA, et al. Immunotherapy with vaccines combining MHC class II/CD80+ tumor cells with interleukin-12 reduces established metastatic disease and stimulates immune effectors and monokine induced by interferon gamma. Cancer Immunol. Immunother. 49: 34-45, 2000. PubMed: 10782864

Pulaski BA, Ostrand-Rosenberg S. Reduction of established spontaneous mammary carcinoma metastases following immunotherapy with major histocompatibility complex class II and B7.1 cell-based tumor vaccines. Cancer Res. 58: 1486-1493, 1998. PubMed: 9537252

Aslakson CJ, Miller FR. Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor. Cancer Res. 52: 1399-1405, 1992. PubMed: 1540948

Pulaski BA, et al. Cooperativity of Staphylococcal aureus enterotoxin B superantigen, major histocompatibility complex class II, and CD80 for immunotherapy of advanced spontaneous metastases in a clinically relevant postoperative mouse breast cancer model. Cancer Res. 60: 2710-2715, 2000. PubMed: 10825145

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