UPCI:SCC090

The patient had a recurrence about 1 year later sited at the hypopharynx. It is ATCC^® No. CRL-3240 UPCI:SCC152™.

These 2 HNSCC (head and neck squamous cell carcinoma) cell lines are a resource for studying the initiation, cancerization, prognosis, intervention and treatment of oral cancers.

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

• additional 2mM L-Glutamine
• 10 mM NEAA
• fetal bovine serum (FBS) to a final concentration of 10%

1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC® No. 30-2200) or 0.25% Trypsin – 0.53mM EDTA (ATCC® No. 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.
2. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
4. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C.
Subcultivation Ratio: 1:2 to 1:4 is recommended.
Medium Renewal: 2 to 3 times a week

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