Description: Derived in 1979 from BALB/c spleen cells activated by sheep erythrocytes in the presence of IL-2, the A5E clone was selected to provide a homogenous population that is highly sensitive to IL-2. This cell line expresses Thy-1 and Lyt markers in addition to high affinity IL-2 receptors. Substantial proliferation is seen in the presence of IL-4 (GM-CSF).

Also Known As:

Species: Mouse

Tissue: Blood

Growth Properties: Suspension

Morphology: Lymphoblast

Growth Medium: RPMI 1640 + 2mM Glutamine + 0.05mM 2-Mercaptoethanol (2ME) + 100-200 IU/ml IL-2 + 10% Foetal Bovine Serum (FBS).

Subculturing Procedure: Maintain cultures between 3-9×100,000 cells/ml; 5% CO2; 37°C. When starting from a frozen ampoule, add thawed cells to a conical based centrifuge tube e.g. 15ml tube. Slowly add 4 ml of culture medium to the tube. Take a sample of the cell suspension, e.g. 200µl, to count the cells. Centrifuge the cell suspension at low speed i.e. 100 – 150 x g for a maximum of 5 minutes. Remove the medium and re-suspend the cell pellet at a density of 3-5 x 100,000 cells/ml in fresh medium containing 20% serum. Incubate flask at 37°C; 5% CO2 and check daily. Keep flask in a vertical position until the cells reach the exponential phase of growth; this can take up to 10 days. Once the culture is established the serum concentration can be reduced to 10%. To keep the cells in exponential growth, maintain cultures between 3-9×100,000 cells/ml. Once the culture is established it can demonstrate rapid growth and may quickly exhaust the medium components and lose viability as a result. We recommend the culture is monitored by cell counting for its growth rate. Fresh IL-2 should be added at each sub-culture.

Release Conditions: No

Depositor:

Originator: No

References: J Immunol 1987;138:4280; Proc Natl Acad Sci, USA 1986;83:2061; J Exp Med 1979;150:1510

Hyperlink to ECACC Cell Line Data Sheet: 92021401