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.
This cell line can be used to study the role of caspase-8 in apoptotic signaling pathways in the absence of overexpression.
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. Ref
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
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.
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.
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).
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
