UPCI:SCC152 is from a squamous cell carcinoma of the hypopharynx. It was a recurrent tumor. The patient’s original tumor site was the base of the tongue. The UPCI:SCC152 cells are positive for Human Papilloma Virus (HPV). These cells have no TP53 mutations as assayed by sequencing the 5-8 exons of TP53. UPCI:SCC152 showed no amplification of chromosomal band 11q13 using FISH.

The base medium for this cell line is ATCC-formulated Eagles Minimum Essential Medium (ATCC^® 30-2003™). To make the complete growth medium, add the following components to the base medium:xa0

• Additional 2 mM L-Glutaminexa0
• Fetal bovine serum (FBS; ATCC^® 30-2020™) to a final concentration of 10%xa0
• Gentamicin to a final concentration of 50 µg/mL
• 5.6 mL NEAA (Gibco Cat# 11440-076)

Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos 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.

The flask was seeded with cells (see specific batch information) grown and completely filled with medium at ATCC to prevent loss of cells during shipping.

1. Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with phase-contrast optics), carefully check for any evidence of microbial contamination.xa0 Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
2. If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium.xa0 The shipping medium can be saved for reuse.xa0 Incubate the cells at 37°C in a 5% CO_2xa0 xa0in air atmosphere until they are ready to be subcultured.
3. If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 125 x g for 5 to 10 minutes.xa0 Remove shipping medium and save.xa0 Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm² flask.xa0 Incubate at 37°C in a 5% CO₂ xa0in air atmosphere until cells are ready to be subcultured.

White JS, et al. The influence of clinical and demographic risk factors in the establishment of head and neck squamous cell carcinoma cell lines. Oral Oncol. 43:701-712, 2007. PubMed: 17112776

Martin CL, et al. Chromosomal imbalances in oral squamous cell carcinoma: examination of 31 cell lines and review of the literature. Oral Oncol. 44:369-382, 2008. PubMed: 17681875