Cell Line Origin
Buccal mucosa squamous carcinoma
Cell Line Description
Adherent keratinocyte cell line derived from a recurrent squamous cell carcinoma of the buccal mucosa of a Caucasian female. Keratin and involucrin markers present. Known mutations: p53, p16 and p14ARF. Cells are tumourigenic in athymic mice.
Study of genotype/phenotype relationships
DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS) + 0.4 micrograms/ml Hydrocortisone.
Split sub-confluent cultures (70-80%) 1:10 - 1:50 using 0.25% trypsin/EDTA; 8% CO2; 37 °C. Suggested seeding density 2 x 10,000 cells/cm2.
Culture cells on a feeder layer of lethally-irradiated or mitomycin C-treated 3T3 Swiss Albino cells (Sigma catalog number 85022108). Feeder layers are prepared in the flasks at least 24 hours in advance of being required. Where frozen stocks of treated 3T3 Swiss Albino cells have been prepared, an ampoule is thawed in 37 °C water bath and the contents quickly transferred to a 15 ml centrifuge tube. DMEM medium is added drop wise to 5 ml. Cells are centrifuged at 150 x g for 5 min at room temperature. Cells are resuspended in 5 ml of medium. Cells are counted and added to flasks containing the correct BICR growth medium at 1-3 x 104 cells/cm2.
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This cell line has special release conditions: Commercial organisations are required to complete the ′Cell Line Release Authorisation for Research Use in Commercial Organisations′ release conditions form.