U-87 MG-Luc2 (ATCC® HTB-14-LUC2)

Organism: Homo sapiens, human  /  Tissue: brain  /  Disease: glioma

Permits and Restrictions

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Organism Homo sapiens, human
Tissue brain
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 glioma
Age 44 years
Gender male
Ethnicity unknown
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 Nu/Nu mice
Comments

This luciferase expressing cell line was derived from parental line HTB-14 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 Eagle's Minimum Essential Medium (EMEM; ATCC 30-2003). 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
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 1 x 104 viable cells/cm2.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2 X 104 and 1 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:5 is recommended
Medium Renewal: 2 to 3 times per week
Maintenance

The flask was seeded with cells (see specific batch information) grown and completely filled with medium at ATCC to prevent loss of cells during shipping.

  1. Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with phase-contrast optics), carefully check for any evidence of microbial contamination.  Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
  2. If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium.  The shipping medium can be saved for reuse.  Incubate the cells at 37°C in a 5% CO in air atmosphere until they are ready to be subcultured.
  3. If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 125 x g for 5 to 10 minutes.  Remove shipping medium and save.  Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask.  Incubate at 37°C in a 5% CO2  in air atmosphere until cells are ready to be subcultured.
Cryopreservation Complete growth medium supplemented with 5% (v/v) DMSO
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 1.0 x 106
Volume 1.0 mL
STR Profile
Amelogenin: X
CSF1PO: 10,11
D13S317: 8,11
D16S539: 12
D5S818: 11,12
D7S820: 8,9
THO1: 9.3
TPOX: 8
vWA: 15,17
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Cytomegalovirus: None detected
Human immunodeficiency virus: None detected
Epstein-Barr virus: None detected
Human papillomavirus: None detected
Functional Tests Luciferase activity: signal to noise ≥ 1,000
In Vitro Luminesence: ≥ 300,000 photons/cell/sec, subject to imaging and culturing conditions.
Name of Depositor ATCC
Year of Origin 2017
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

Allen M, et al. Origin of the U87MG glioma cell line: Good news and bad news. Sci. Trans. Med. 8(354): 1-4, 2016.

Clark MJ, et al. U87MG decoded: The genomic sequence of a cytogenetically aberrant human cancer cell line. PLoS Genetics 6 (1) : e1000832, 2010.

Lieber M, et al. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer 17: 62-70, 1976. PubMed: 175022

Lewis JA, et al. Inhibition of mitochondrial function by interferon. J. Biol. Chem. 271: 13184-13190, 1996. PubMed: 8662694

Jamaluddin M, et al. Inducible translational regulation of the NF-IL6 transcription factor by respiratory syncytial virus infection in pulmonary epithelial cells. J. Virol. 70: 1554-1563, 1996. PubMed: 8627674

Zinn K, et al. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell 34: 865-879, 1983. PubMed: 6313211

Li YM, et al. Molecular identity and cellular distribution of advanced glycation endproduct receptors: relationship of p60 to OST-48 and p90 to 80K-H membrane proteins. Proc. Natl. Acad. Sci. USA 93: 11047-11052, 1996. PubMed: 8855306

Olopade OI, et al. Molecular analysis of deletions of the short arm of chromosome 9 in human gliomas. Cancer Res. 52: 2523-2529, 1992. PubMed: 1568221

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Beckman G, et al. G-6-PD and PGM phenotypes of 16 continuous human tumor cell lines. Evidence against cross-contamination and contamination by HeLa cells. Hum. Hered. 21: 238-241, 1971. PubMed: 4332744

Ponten J, Macintyre EH. Long term culture of normal and neoplastic human glia. Acta Pathol. Microbiol. Scand. 74: 465-486, 1968. PubMed: 4313504

Tumors developed within 21 days at 100% frequency (5/5).

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement: Luciferase Label License.
For information on obtaining additional rights, please contact:
ATCC Licensing
Email: licensing@atcc.org

For commercial accounts, this cell line is only distributed under the terms of a fully signed and executed ATCC® Material Transfer Agreement and Addendum. If the commercial account is screening per completed Addendum, the recipient will be required to pay a Screening Fee (ATCC® ACS-2103F™).

Screening Use is defined as use of Biological Material in small molecule and biologic drug discovery, including initial target identification and validation, assay development, high throughput screening, hit identification, lead optimization, and selection of candidates for clinical development.

If the commercial account is not screening per the completed Addendum, the recipient will not be required to pay a Screening Fee.

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

Allen M, et al. Origin of the U87MG glioma cell line: Good news and bad news. Sci. Trans. Med. 8(354): 1-4, 2016.

Clark MJ, et al. U87MG decoded: The genomic sequence of a cytogenetically aberrant human cancer cell line. PLoS Genetics 6 (1) : e1000832, 2010.

Lieber M, et al. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer 17: 62-70, 1976. PubMed: 175022

Lewis JA, et al. Inhibition of mitochondrial function by interferon. J. Biol. Chem. 271: 13184-13190, 1996. PubMed: 8662694

Jamaluddin M, et al. Inducible translational regulation of the NF-IL6 transcription factor by respiratory syncytial virus infection in pulmonary epithelial cells. J. Virol. 70: 1554-1563, 1996. PubMed: 8627674

Zinn K, et al. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell 34: 865-879, 1983. PubMed: 6313211

Li YM, et al. Molecular identity and cellular distribution of advanced glycation endproduct receptors: relationship of p60 to OST-48 and p90 to 80K-H membrane proteins. Proc. Natl. Acad. Sci. USA 93: 11047-11052, 1996. PubMed: 8855306

Olopade OI, et al. Molecular analysis of deletions of the short arm of chromosome 9 in human gliomas. Cancer Res. 52: 2523-2529, 1992. PubMed: 1568221

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Beckman G, et al. G-6-PD and PGM phenotypes of 16 continuous human tumor cell lines. Evidence against cross-contamination and contamination by HeLa cells. Hum. Hered. 21: 238-241, 1971. PubMed: 4332744

Ponten J, Macintyre EH. Long term culture of normal and neoplastic human glia. Acta Pathol. Microbiol. Scand. 74: 465-486, 1968. PubMed: 4313504

Tumors developed within 21 days at 100% frequency (5/5).