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BCRJ Code 0250
Cell Line XR63-IL2
Species Mus musculus
Vulgar Name Mouse
Tissue Hematopoietic
Cell Type Myeloid
Morphology Lymphoblast
Disease Myeloma
Growth Properties Suspension
Derivation XR63-IL2 cell line was derived from X63Ag-654 myeloma cell line.
Products Interleukin-2; IL-2
Biosafety 1
Addtional Info It was transformed by pBV-1MTHA vecctor containing IL-2 ccDNA cloned by D W. Fiers (State University of Ghent, Belgium). cells stably carry 30-100 copies of the plasmid per cell and constitutively secrete biolocally active mouse IL-2 in quantities similar to EL-4 thymoma cells and rat spleen cells stimulated with mitogens.
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 1 mM sodium pyruvate, 4500 mg/L glucose and 10% of fetal bovine serum.
Subculturing Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 3 X 10e5 cells/mL and maintain between 2 X 10e5 and 10e6 cells/mL.
Subculturing Medium Renewal Every 2 to 3 days
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 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).
References Eur. J. Immunol. 18: 97-104, 1988.
Depositors Dr. Fritz Melchers , Basel Institute for Immunology, Basel, Switzerland.