Established HNSCC Cell Lines for Research

Within this study we worked with several established human squamous cell carcinoma (SCC) lines of the head and neck. The FaDu_DD [37 (link)] cells originate from the hypopharynx; SAS (JCRB Cell Bank, NIBIOHN, Osaka, Japan), Cal33 (DSMZ, Braunschweig, Germany) and UT-SCC-5 cells (DSMZ) from the tongue. The metastatic cell line Detroit562 (CLS, Eppelheim, Germany) derives from pleural effusion of a primary tumor of the pharynx. The keratinocyte cell line HaCaT (DKFZ, Heidelberg, Germany) was used as normal tissue control. These established HNSCC cell lines were cultivated as monolayer with Dulbecco’s modified Eagle medium (DMEM; Biochrom, Cambridge, UK) containing 10% fetal calf serum (FCS, Sigma-Aldrich, St. Louis, MO, USA), 2% HEPES buffer (1 M, Biochrom), 1% non-essential amino acids (NEA 100×, Biochrom), 1% sodium pyruvate (Biochrom), and 1% penicillin/streptomycin (10,000 U/mL, Biochrom). The incubator was adjusted to 37 °C and 5% CO₂. Cells were passaged usually twice per week after a confluency of 70–80% was reached. Cells were only used for experiments until passage 15 and regularly tested for cell authentication and mycoplasma infection. Irradiated (IR) sublines were generated from indicated parental cell line by selection with at least 15 fractions of 4 Gy and analyzed together with age-matched controls [36 (link)].