IDH2 mutant-TF-1 Isogenic Cell Line

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.

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

• 2 ng/ml recombinant human GM-CSF
• Fetal Bovine Serum (FBS; ATCC 30-2020) to a final concentration of 10%

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 150 x g for 5 to 7 minutes.
4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm² or a 75 cm² culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

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