||Jurkat, Clone E6-1
||Acute T Cell Leukemia
||The Jurkat cell line was established from the peripheral blood of a 14 year old boy by Schneider et al., and was originally designated JM.
||This cell line is a suitable transfection host.
||Interleukin 2 (IL-2), human alpha interferon
||Clone E6-1 cells produce large amounts of IL-2 after stimulation with phorbol esters and either lectins or monoclonal antibodies against the T3 antigen (both types of stimulants are needed to induce IL-2 production.
||RPMI 1640 medium with 2 mM L-glutamine, 4.5 g/L glucosen and 10% of heat-inactivated fetal bovine serum.
||Cultures can be maintained by the addition of fresh medium or replacement of medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL.
NOTE: Do not allow the cell density to exceed 3 X 10e6 cells/mL.
|Subculturing Medium Renewal
||2 to 3 days
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||95% FBS + 5% DMSO (Dimethyl sulfoxide)
|Thawing Frozen Cells
||SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that
some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line).
NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
||J. Exp. Med. 152: 1709-19, 1980;
J. Immunol. 133: 123-128, 1984.
Berninghausen O, Leippe M. Necrosis versus apoptosis as the mechanism of target cell death induced by Entamoeba histolytica. Infect. Immun. 65: 3615-3621, 1997. PubMed: 9284127
Churchill MJ, et al. The rev-responsive element negatively regulates human immunodeficiency virus type 1 env mRNA expression in primate cells. J. Virol. 70: 5786-5790, 1996. PubMed: 8709194
Kolanus W, et al. alphaLbeta2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1 a cytoplasmic regulatory molecule. Cell 86: 233-242, 1996. PubMed: 8706128
||Pedro Paulo Elssas, Instituto Oswaldo Cruz.