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IDH2 mutant-TF-1 Isogenic-Luc2 (ATCC® CRL-2003IG-LUC2)

Organism: Homo sapiens, human  /  Cell Type: erythroblast  /  Tissue: bone marrow  /  Disease: erythroleukemia

Permits and Restrictions

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Organism Homo sapiens, human
Tissue bone marrow
Cell Type erythroblast
Product Format frozen 1.0 mL
Morphology lymphoblast
Culture Properties suspension
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 erythroleukemia
Age 35 years
Gender male
Ethnicity Japanese
Applications IDH2 mutant leukemia model. 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 NSG mice
Comments This luciferase expressing cell line was derived from parental line ATCC CRL-2003IG by transduction with lentiviral vector encoding firefly luciferase gene (luc2) under control of EF-1 alpha promoter. The parental line CRL-2003IG is a CRISPR gene edited leukemia model containing IDH2 R140Q mutations. 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:
  • 2 ng/ml recombinant human GM-CSF
  • Fetal Bovine Serum (FBS; ATCC 30-2020) to a final concentration of 10%
  • Blasticidin to a final concentration of 8µg/mL
Subculturing
Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 8 X 104 cells/mL and maintain between 5 X 104 and 1 X 106 cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density).
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 cells
Volume 1.0 mL
STR Profile
Amelogenin: X,Y
CSF1PO: 13
D13S317: 8,9
D16S539: 9,12
D5S818: 13
D7S820: 12
TH01: 8
TPOX: 7,9
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 RLUs
in vitro Luminesence: 100,000 photons/cell/sec, subject to imaging and culturing conditions
Population Doubling Time approximately 30 hrs
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

Kroeze LI, et al. Characterization of acute myeloid leukemia based on levels of global hydroxymethylation. Blood 124(7): 1110-1118, 2014. PubMed: 24986689

Wang F, et al. Targeted inhibition of mutant IDH2 in leukemia cells induces cellular differentiation. Science 340(6132): 622-626, 2013. PubMed: 23558173

Ward PS, et al. The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate. Canc Cell 18(3): 225-234, 2010. PubMed: 20171147

Figueroa ME, et al. Leukemic IDH1 and IDH2 mutations result in a hypermethylation phenotype, disrupt TET2 function, and impair hematopoietic differentiation. Canc Cell 18(6): 553-567, 2010. PubMed: 21130701

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.

In addition to the foregoing, this product's use is governed by the CRISPR Label License Agreement. For information on purchasing a license to use this product for purposes other than those permitted in the CRISPR Label License Agreement, please contact The Broad Institute at partnering@broadinstitute.org.

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. CRISPR Label License, ERS Genomics

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: licensing@atcc.org

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

Kroeze LI, et al. Characterization of acute myeloid leukemia based on levels of global hydroxymethylation. Blood 124(7): 1110-1118, 2014. PubMed: 24986689

Wang F, et al. Targeted inhibition of mutant IDH2 in leukemia cells induces cellular differentiation. Science 340(6132): 622-626, 2013. PubMed: 23558173

Ward PS, et al. The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate. Canc Cell 18(3): 225-234, 2010. PubMed: 20171147

Figueroa ME, et al. Leukemic IDH1 and IDH2 mutations result in a hypermethylation phenotype, disrupt TET2 function, and impair hematopoietic differentiation. Canc Cell 18(6): 553-567, 2010. PubMed: 21130701