I 9.2 (ATCC® CRL-2571)

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 caspase-8 in apoptotic signaling pathways in the absence of overexpression.

Storage Conditions liquid nitrogen vapor phase
Derivation

The I 9.2 cell line is a caspase-8 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

The I 9.2 cell line is functionally defective for caspase-8 and the I 2.1 cell line is functionally defective for FADD. The I 9.2 cell line and the I 2.1 cell lines are completely resistant to Fas-induced apoptosis. 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 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 

Complementation of I 9.2 cell line with wild-type caspase-8 restored Fas-mediated apoptosis. 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   

The I 9.2 cell line is severely deficient in cell death induced by TNF-alpha and is partially deficient in cell death induced by ultraviolet irradiation, adriamycin and etoposide. 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 This cell line can be used to study the role of caspase-8 in apoptotic signaling pathways in the absence of overexpression.


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 10%.
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 cultures at cell concentrations between 2 x 105 and 2 x 106 viable cells/mL. Do not allow the cell concentration to exceed 2 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. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 11,12
D13S317: 8,11
D16S539: 11
D5S818: 9
D7S820: 8,9.2
THO1: 6,9.3
TPOX: 8,10
vWA: 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

Ding HF, et al. Essential role for caspase-8 in transcription-independent apoptosis triggered by p53. J. Biol. Chem. 275: 38905-38911, 2000. PubMed: 10988287

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

Ding HF, et al. Essential role for caspase-8 in transcription-independent apoptosis triggered by p53. J. Biol. Chem. 275: 38905-38911, 2000. PubMed: 10988287