Description: The T cell leukaemia line J.CaM1.6 was derived from the parent cell line Jurkat E.6-1 (ECACC catalogue no. 88042803). Jurkat cells were mutagenised with ethyl methanesulfonate and subsequently grown in phytohemagglutinin containing media for 2 weeks. J.CaM1.6 express a structurally normal but functionally defective CD3/Ti complex. The cells fail to exhibit inositolphospholipid metabolism or Ca²+ mobilisation in response to anti-CD3 or anti-Ti monoclonal antibodies. They display resistance to anti-CD3 induced apoptosis, constitutively express Fas and are sensitive to anti-Fas induced apoptosis. It was shown that J.CaM1.6 cells are defective in early signalling events that follow TCR stimulation owing to loss of lck tyrosine kinase function due to the lack of exon 7 in the lck transcript.

Also Known As:

Species: Human

Tissue: blood

Growth Properties: Suspension

Morphology: Lymphoblast

Growth Medium: RPMI 1640 + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).

Subculturing Procedure: To keep the cells in exponential growth, maintain cultures between 3-9×100,000 cells/ml; 5% CO2; 37°C. If 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. 100ul, to count cells. Centrifuge the cell suspension at low speed i.e. 100 – 150 x g for a maximum of 5 minutes. Remove medium and re-suspend the cell pellet at a density of 3-5 x 100,000 cells/ml in fresh medium containing 10% serum. Incubate flask at 37°C; 5 – 7% CO2. Check daily. Keep flask in a vertical position until the cells reach the exponential phase of growth, this can take up to 7 days. .

Release Conditions: No

Depositor:

Originator: No

References: Proc Natl Acad Sci, USA 1987;84:6879; Cell 1992;70:585; Biochem Biophys Res Comm 1995;213:994

Hyperlink to ECACC Cell Line Data Sheet: 96060401