ATCC Gold Seal IDH2 mutant-TF-1 Isogenic Cell Line (ATCC® CRL-2003IG)

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

Permits and Restrictions

View Permits View Restrictions

Organism Homo sapiens, human
Tissue bone marrow
Verified By ATCC Cell Line Authentication Service
Functional Testing
Sanger Sequencing
Cell Type erythroblast
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 2  [Cells containing CMV and SV40 viral sequences]

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 Tumor-associated mutant IDH2R140Q, isocitrate dehydrogenase-2, IDH2 mutant, acute myeloid leukemia (AML) patients, anti-cancer drug screening, IDH2-specific inhibitors.
Storage Conditions liquid nitrogen vapor phase
Images CRL-2003IG Validation of IDH2 inibitors CRL-2003IG Validation of IDH2 inibitors decrease 2-HG levels ATCC CRL-2003IG, IDH2 mutant-TF-1 Isogenic Cell Micrograph
Comments

This is an acute myeloid leukemia (AML) IDH2R140Q mutant isogenic line derived from the parental TF-1 cell line. The c.419G>A knock-in mutation encoding IDH2R140Q protein expression was generated at ATCC by utilizing the CRISPR/Cas9 gene editing technology. This is a homozygous mutation expressing the c.419G>A mutant allele. The IDH2R140Q mutation in CRL-2003IG has been validated at the genomic, transcript, and protein bio-functional levels.

The IDH2R140Q mutation is a cancer driver gene which causes a gain of function in tumor cells converting isocitrate to the oncometabolite, D-2-hydroxyglutarate (D-2HG). High intracellular levels of D-2HG inhibits alpha-ketoglutarate-dependent DNA and histone demethylases, resulting in epigenetic disregulation, which in turn leads to a block in cellular differentiation, promoting AML development. These effects are reduced with treatment of IDH2 mutant-specific small-molecule inhibitors such as AG-221.

The IDH2R140Q mutant isogenic line CRL-2003IG has been tested for neomorphic functional activity displaying elevated intra- and extracellular D-2HG levels. A reduction in histone hypermethylation was also observed upon treatment with IDH2 specific molecules, AG-221 and AGI-6780. This IDH2R140Q isogenic cell model is a valuable in vitro cell-based tool for clinical diagnostics, elucidating mechanisms involved in cancer-associated differentiation, tumorigenesis and use in screening anti-cancer compounds for drug discovery and development.

Refer to the TF-1 parental line (ATCC CRL-2003) for additional background information.

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%
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).
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 1 x 106 cells
STR Profile
Amelogenin: X,Y
CSF1PO: 13
D13S317: 8,9
D16S539: 9,12
D5S818: 13
D7S820: 12
THO1: 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 interleukin 3 (IL-3) independent proliferation assay; intracellular and extracelluylar D-2-hydroxyglutarate (D-2HG) measurement, histone hypermethylation levels
Population Doubling Time approximately 24.8 hours
Year of Origin 2016
References

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
FAQ's
  1. IDH2 mutant-TF-1 Isogenic Cell Line ATCC CRL-2003IG


    Date Updated: 10/3/2017

  2. Validation - CRL-2003IG


    Date Updated: 10/3/2017

  3. Applications of the IDH2 mutant-TF-1 Isogenic Cell Line


    Date Updated: 10/3/2017

Restrictions

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.

References

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