Bjhtert

HEK293T, U20S, and BJhTERT cell lines were obtained from ATCC and grown in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS), 1X Penicillin/Streptomycin (Pen Strep, catalog # 15140122). Merkel Cell Carcinoma cell line MKL-1 was obtained from ECACC (# 09111801) and MS-1, MCC26, and WaGa cells were kind gifts from Dr. James Decaprio's laboratory at Dana Farber Cancer Institute, Boston. MCC cell lines were cultured in Roswell Park Memorial Institute (RPMI) Media with 10% FBS and 1X Penicillin/Streptomycin. The cells were grown at 37°C with 5% CO₂.

PC3, MDA-MB-231, MCF7, HEK-293T, MCF10A, and BJ-hTert cells used in this study were obtained from the American Type Tissue Culture Collection. Mouse PNET N134 cells were generated by the Du laboratory[29 (link)]. PC3 cells were maintained in F-12K medium, MCF10A cells were maintained in MEGM Mammary Epithelial Cell Growth Medium Bullet Kit (Lonza) supplemented with 100 ng/ml cholera toxin. other cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM), supplemented with 10%, or 15% (N134) fetal bovine serum (Hyclone). No further authentication was performed.

HCT116, MDA-MB-231, MCF7, and BJ-hTert cells used in this study were obtained from the American Type Tissue Culture Collection. HCT116 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (Sigma D5796) supplemented with 10% Fetal Bovine Serum (FBS) (Sigma 12306C). All other cells were maintained in DMEM (Sigma D6429) supplemented with 10% FBS (Sigma 12306C). Media used for glutamine free conditions were DMEM (Sigma D5671) for HCT116 cells and DMEM (Sigma D5546) for all other cell lines. All culture media was supplemented with 1x antibiotic/antimycotic solution (Sigma A5955). No further authentication was performed.

The Human pancreatic cancer cell lines PANC1, ASPC1 and the hTERT-immortalized foreskin fibroblast BJhTERT were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). PANC28 cell line was obtained from the laboratory of Dr. Marsha L. Fraizer and Dr. Douglas B. Evans. In adherent condition all cell lines were maintained as monolayer cultures and cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 4.5 g/L glucose, glutamine, and non-essential amino acids and supplemented with 10% heat-inactivated fetal bovine serum and penicillin (100 IU/mL)–streptomycin (100 μg/mL). In spheroid-forming condition all cell lines were plated (4000 cells/ml) in low attachment plates and cultured 48 h in sphere medium (DMEM / F12 supplemented with BSA 0.1%, glucose 0.5%, heparin 4 μg/ml, L-glutamine 2.5 mM, PS 1X, FGF 20 ng/ml, EGF 20 ng/ml, B27 1X, insulin 20 μg/ml) to perform the assays shown. Cultures were maintained in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. All cell lines were regularly inspected for mycoplasma. The cells have been authenticated with short tandem repeat profile generated by LGC Standards.

Head and Neck squamous cancer carcinoma cell lines FaDu, SCC9, and Normal fibroblasts BJ-hTERT, were purchased from the American Type Culture Collection (ATCC, Rockville, MD, United States); Cal27 cell lines were kindly provided by Dr. J.L. Fishel (Centre A Lacassagne, Nice, France). The green fluorescent protein⁺/luciferase⁺ (GFP⁺/Luc⁺) Cal27 cell line were obtained by lentiviral infection as described previously (Piro et al., 2019 (link)). HOC313 and ZA cell lines were kindly provided by Dr. N. Tsuchida (Faculty of Medicine, Saitama Medical University, Saitama, Japan) (Tadokoro et al., 1989 (link)).
Cal27, ZA, SCC9, HOC313, and BJ-hTERT cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM), whereas FaDu were cultured in RPMI-1640 medium. All media were supplemented with 10% heat-inactivated fetal bovine serum (for ZA cells media was supplemented with 20% of heat-inactivated fetal bovine serum), 50 units/mL penicillin, 500 μg/mL streptomycin, and 4 mmol/L glutamine. Cultures were maintained in a humidified atmosphere of 95% air and 5% CO₂ at 37°C.
All cell lines were regularly inspected for mycoplasma. The cells have been authenticated with short tandem repeat profile generated by LGC Standards.