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CRL-2572 A3 人T**细胞白血病细胞
 

ATCC® Number: CRL-2572™     Price:  
Designations: I 2.1
Depositors:  J Blenis
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: suspension
Organism: Homo sapiens (human)
Morphology: lymphoblast
Source: Disease: acute T cell leukemia 
Cell Type: T lymphocyte;
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
 
DNA Profile (STR): 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
Gender: male
Comments: 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. [51527] 
ICR-191 treated clones were serially diluted in 96-well plates in the presence of Fas Antibody for 3 to 5 weeks. [51529] 
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. 
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. [51529] 
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. [51529] 
This cell line can be used to study the role of FADD in apoptotic signaling pathways in the absence of overxpression.
Propagation: ATCC 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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5% 
Temperature: 37.0°C
Subculturing: Protocol: 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 10(5) viable cells/ml. Maintain cell density between 2 X 10(5) and 2 X 10(6) viable cells/ml. 
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO 
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
parental cell line:ATCC CRL-2570
References: 51527: 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
51529: 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
51530: 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

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