Isolation of OSCC Cell Lines

Cell lines were isolated from patients with oral squamous cell carcinoma (OSCC) as mentioned in previous work [9 (link)]. Briefly, tumors were minced and enzymatically dissociated using 4 mg/mL-1 Collagenase type IV (Thermo Fisher, cat. no. 17104019) in DMEM/F12, at 37 °C for 1 h. Post digestion, cells were pelleted and resuspended in phosphate-buffered saline (Thermo Fisher, cat. no 14190235) for 3 cycles. Cells were then strained through 70-μm cell strainers (Falcon, cat. no. 352350), prior to pelleting and resuspension in RPMI media (Thermo Fisher, cat. no 61870036), containing 10% fetal bovine serum (Gibco, cat. no 10270-106) and 1% penicillin-streptomycin (Thermo Fisher, cat. no. 15140122). Cells were plated on CellBIND plates (Corning, cat. no 3335) and kept in a humidified atmosphere of 5% CO₂ at 37 °C. Cells were routinely screened for mycoplasma contamination using MycoAlertTM PLUS Mycoplasma Detection Kit (Lonza, cat. no: LT07-710).