ATCC Number：TIB-214?
品系：C57BL/6
細胞形態(tài)：淋巴樣
細胞類型：其他細胞類型
是否是腫瘤細胞：0
物種來源：小鼠
生長狀態(tài)：懸浮生長
數(shù)量：大量
運輸方式：凍存運輸
規(guī)格：0.1ml Designations: CTLL-2
Depositors: ?S Gillis
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:suspension
Organism: Mus musculus
Morphology:lymphoblast
Source: Cell Type: lymphocyte cytotoxic T lymphocyte;
Strain: C57BL/6
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.
Applications:The freshly prepared rat factor will usually produce more rapid growth.
This line is a clone of cytotoxic T cells derived from a C57BL/6 mouse.
The cells are dependent upon IL-2 for growth, and can be used to assay for IL-2. Immediately after thawing (or after overgrowth of a culture) the culture will appear to have few or no viable cells.
T-STIM with Con A (rat IL-2 culture supplement from Becton Dickinson) may be used or the rat factor may be prepared as described on the product sheet.
Tested and found negative for ectromelia virus (mousepox).
Receptors:interleukin 2 (IL-2), expressed
Comments:This line is a clone of cytotoxic T cells derived from a C57BL/6 mouse. The cells are dependent upon IL-2 for growth, and can be used to assay for IL-2. Immediately after thawing (or after overgrowth of a culture) the culture will appear to have few or no viable cells. This is normal. Depending upon the source and potency of the IL-2, it may take up to three weeks before the cells are ready to subculture. T-STIM with Con A (rat IL-2 culture supplement from Becton Dickinson) may be used or the rat factor may be prepared as described on the product sheet. The freshly prepared rat factor will usually produce more rapid growth. Tested and found negative for ectromelia virus (mousepox).
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: additional 2 mM L-glutamine; additional 1mM sodium pyruvate; adjust to a final concentration of 10% fetal bovine serum and 10% T-STIM with Con A. T-STIM is available from Becton Dickinson.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0℃
Subculturing: Protocol: Some Important Considerations in Handling CTLL-2, TIB-214:Frozen Cells: Viability immediately after thawing will be 70-80%. Expect viability to be very poor from day 1 to day 4 after culture initiation. Culture will appear to be completely dead. On the third to fifth day following initiation viable cell clusters will begin to appear in suspension. Usually cells will be ready to subculture on the 7th to the 10th day after the ampule is thawed. However, it may take from 2-3 weeks before vigorous growth is observed. It is best to leave the initial culture undisturbed until cells enter their growth phase.
Overgrowth: In the event cell density becomes too great and viability decreases to where culture appears totally dead, the culture may still be rescued. Inoculate a flask at a density of 1 X 10(4) viable cells/ml.
Subculturing Procedure:Subculture actively growing suspension cultures before they have reached 2 X 10(5) cells/ml or the IL-2 will rapidly deplete and the cells will quickly lose viability Use inoculation densities of 1 to 2 X 10(4) viable cells/ml.
Medium Renewal: Twice per week
Preservation: Freeze medium: Complete growth medium supplemented with an additional 10% fetal bovine serum and 7.5% 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
References: 22901: Gillis S, Smith KA. Long term culture of tumour-specific cytotoxic T cells. Nature 268: 154-156, 1977. PubMed: 145543
32283: Hu SX, et al. Development of an adenovirus vector with tetracycline-regulatable human tumor necrosis factor alpha gene expression. Cancer Res. 57: 3339-3343, 1997. PubMed: 9269991
32343: Mazzaccaro RJ, et al. Major histocompatibility class I presentation of soluble antigen facilitated by Mycobacterium tuberculosis infection. Proc. Natl. Acad. Sci. USA 93: 11786-11791, 1996. PubMed: 8876215
49841: Belani R, Weiner GJ. Expression of both B7-1 and CD28 contributes to the IL-2 responsiveness of CTLL-2 cells. Immunology 87: 271-274, 1996. PubMed: 8698390
