I 2.1 (ATCC® CRL-2572)

Organism: Homo sapiens, human  /  Cell Type: T lymphocyte  /  Disease: acute T cell leukemia

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Organism Homo sapiens, human
Cell Type T lymphocyte
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Disease acute T cell leukemia
Gender male
Applications

This cell line can be used to study the role of FADD in apoptotic signaling pathways in the absence of overxpression.

Storage Conditions liquid nitrogen vapor phase
Derivation The I 2.1 cell line is a FADD mutant of the wild-type Jurkat cell line A3 (see ATCC -CRL-2570). Wild-type A3 cells were made neomycin resistant and treated with three cycles of exposure to the frameshifting mutagen ICR-191 to isolate clones harboring recessive mutations that were resistant to killing by Fas antibody. RefJuo P, et al. Essential requirement for caspase-8/FLICE in the initiation of the Fas-induced apoptotic cascade. Curr. Biol. 8: 1001-1008, 1998. PubMed: 9740801

ICR-191 treated clones were serially diluted in 96-well plates in the presence of Fas Antibody for 3 to 5 weeks. RefJuo P, et al. FADD is required for multiple signaling events downstream of the receptor Fas. Cell Growth Differ. 10: 797-804, 1999. PubMed: 10616904 Two of these ICR-191 treated clones have been deposited at the ATCC . They are I 9.2 (ATCC CRL-2571), a clone with a mutation in the cysteine protease caspase-8/FLICE and I 2.1 (ATCC CRL-2572), a clone with a mutation in the adaptor FADD. 

Comments

Unlike the parental line A3 that does express FADD, the I 2.1 cell line does not express FADD protein by immunoblot analysis and is completely resistant to Fas-induced death. Complementation of the I 2.1 cell lines with wild-type FADD restores Fas-mediated apoptosis. Fas activation of caspase-2, caspase-3, caspase-7, and caspase-8 is completely defective in the FADD mutant cell line I 2.1. RefJuo P, et al. FADD is required for multiple signaling events downstream of the receptor Fas. Cell Growth Differ. 10: 797-804, 1999. PubMed: 10616904

This cell line can be used to study the role of FADD in apoptotic signaling pathways in the absence of overxpression. 

Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 15%.
Subculturing
Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension in fresh medium at 3 to 5 x 105 viable cells/mL. Maintain cell density between 2 x 105 and 2 x 106 viable 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. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 8,11
D16S539: 11
D5S818: 9
D7S820: 8,10
THO1: 6,9.3
TPOX: 8,10
vWA: 17,18
Name of Depositor J Blenis
References

Juo P, et al. Essential requirement for caspase-8/FLICE in the initiation of the Fas-induced apoptotic cascade. Curr. Biol. 8: 1001-1008, 1998. PubMed: 9740801

Juo P, et al. FADD is required for multiple signaling events downstream of the receptor Fas. Cell Growth Differ. 10: 797-804, 1999. PubMed: 10616904

Juo P, et al. Fas activation of the p38 mitogen-activated protein kinase signalling pathway requires ICE/CED-3 family proteases. Mol. Cell. Biol. 17: 24-35, 1997. PubMed: 8972182

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
References

Juo P, et al. Essential requirement for caspase-8/FLICE in the initiation of the Fas-induced apoptotic cascade. Curr. Biol. 8: 1001-1008, 1998. PubMed: 9740801

Juo P, et al. FADD is required for multiple signaling events downstream of the receptor Fas. Cell Growth Differ. 10: 797-804, 1999. PubMed: 10616904

Juo P, et al. Fas activation of the p38 mitogen-activated protein kinase signalling pathway requires ICE/CED-3 family proteases. Mol. Cell. Biol. 17: 24-35, 1997. PubMed: 8972182