HT-1080-Luc2 (ATCC® CCL-121-LUC2)

Organism: Homo sapiens, human  /  Tissue: connective tissue  /  Disease: fibrosarcoma

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Organism Homo sapiens, human
Tissue connective tissue
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 fibrosarcoma
Age 35 years
Gender male
Ethnicity Caucasian
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 ATCC CCL-121 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 ATCC-formulated 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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation Complete growth medium supplemented with 5% DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 1.0 x 106 cells
Volume 1.0 mL
STR Profile
Amelogenin: X,Y
CSF1PO: 12
D13S317: 12,14
D16S539: 9,12
D5S818: 11,13
D7S820: 9,10
THO1: 6
TPOX: 8
vWA: 14,19
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: ≥ 100,000 photons/cell/sec, subject to imaging and culturing conditions.
Population Doubling Time approximately 11 hours
Name of Depositor ATCC
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

Chen TR, et al. Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins. Cancer Genet. Cytogenet. 10: 351-362, 1983. PubMed: 6652615

Geiser AG, et al. Suppression of tumorigenicity in human cell hybrids derived from cell lines expressing different activated ras oncogenes. Cancer Res. 49: 1572-1577, 1989. PubMed: 2647289

Rasheed S, et al. Characterization of a newly derived human sarcoma cell line (HT-1080). Cancer 33: 1027-1033, 1974. PubMed: 4132053

Adams RA, et al. Direct implantation and serial transplantation of human acute lymphoblastic leukemia in hamsters, SB-2. Cancer Res. 28: 1121-1125, 1968. PubMed: 4872716

Hu M, et al. Purification and characterization of human lung fibroblast motility-stimulating factor for human soft tissue sarcoma cells: identification as an NH2-terminal fragment of human fibronectin. Cancer Res. 57: 3577-3584, 1997. PubMed: 9270031

Iida A, et al. Inducible gene expression by retrovirus-mediated transfer of a modified tetracycline-regulated system. J. Virol. 70: 6054-6059, 1996. PubMed: 8709228

Brenneman M, et al. Stimulation of intrachromosomal homologous recombination in human cells by electroporation with site-specific endonucleases. Proc. Natl. Acad. Sci. USA 93: 3608-3612, 1996. PubMed: 8622983

Seiffert D. Hydrolysis of platelet vitronectin by calpain. J. Biol. Chem. 271: 11170-11176, 1996. PubMed: 8626663

Hocking AM, et al. Eukaryotic expression of recombinant biglycan. J. Biol. Chem. 271: 19571-19577, 1996. PubMed: 8702651

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’s use is governed by the NanoLuc Limited Use Label License. For information on purchasing a license to use this product for purposes other than those permitted in the Label License, please contact Promega.

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

Chen TR, et al. Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins. Cancer Genet. Cytogenet. 10: 351-362, 1983. PubMed: 6652615

Geiser AG, et al. Suppression of tumorigenicity in human cell hybrids derived from cell lines expressing different activated ras oncogenes. Cancer Res. 49: 1572-1577, 1989. PubMed: 2647289

Rasheed S, et al. Characterization of a newly derived human sarcoma cell line (HT-1080). Cancer 33: 1027-1033, 1974. PubMed: 4132053

Adams RA, et al. Direct implantation and serial transplantation of human acute lymphoblastic leukemia in hamsters, SB-2. Cancer Res. 28: 1121-1125, 1968. PubMed: 4872716

Hu M, et al. Purification and characterization of human lung fibroblast motility-stimulating factor for human soft tissue sarcoma cells: identification as an NH2-terminal fragment of human fibronectin. Cancer Res. 57: 3577-3584, 1997. PubMed: 9270031

Iida A, et al. Inducible gene expression by retrovirus-mediated transfer of a modified tetracycline-regulated system. J. Virol. 70: 6054-6059, 1996. PubMed: 8709228

Brenneman M, et al. Stimulation of intrachromosomal homologous recombination in human cells by electroporation with site-specific endonucleases. Proc. Natl. Acad. Sci. USA 93: 3608-3612, 1996. PubMed: 8622983

Seiffert D. Hydrolysis of platelet vitronectin by calpain. J. Biol. Chem. 271: 11170-11176, 1996. PubMed: 8626663

Hocking AM, et al. Eukaryotic expression of recombinant biglycan. J. Biol. Chem. 271: 19571-19577, 1996. PubMed: 8702651