Nthy ori 3

In this study we used the human non-tumorigenic thyroid cell line Nthy-ori 3.1 (European Collection of Authenticated Cell Culture), which is widely adopted as a model of normal human thyroid cells [38 (link)], and a human PTC cell line, TPC-1, provided by Prof. A Fusco (University of Naples), which is characterized by the RET/PTC1 rearrangement [39 (link)]. Nthy-ori 3.1 and TPC-1 cells were cultured with RPMI 1640 (Thermo Fisher Scientific Inc., Waltham, MA, USA; code 61870) and DMEM (Thermo Fisher Scientific Inc.; code 31966) medium, respectively. The media were supplemented with 10% (v/v) heat-inactivated fetal bovine serum (Thermo Fisher Scientific Inc., Waltham, MA, USA; code 10500064), penicillin (100 IU/mL), streptomycin (0.1 mg/mL) (Thermo Fisher Scientific Inc., Waltham, MA, USA; code 15140-122) and amphotericin B (2.5 µg/mL) (Thermo Fisher Scientific Inc., Waltham, MA, USA; code 30-003). The cells were grown in a monolayer at 37 °C in 5% CO₂ and humidified, as previously described [13 (link)]. To confirm the identity of the cell lines, short tandem repeat analysis was performed by using an AmpFLSTR NGM SElect PCR Amplification Kit (Thermo Fisher Scientific Inc., Waltham, MA, USA; code 4457889).

The thyroid cancer cell lines BC-PAP, TPC-1, 8505c and FTC133, and the normal thyroid cell line NTHY-ORI3-1 were purchased from Gibco (Thermo, Waltham, MA, USA). The thyroid cancer cell lines BC-PAP, TPC1, and 8505c were cultured with Dulbecco’s modified Eagle’s medium (DMEM) (Thermo, Gibco, MA, USA), and supplemented with 10% fetal bovine serum (FBS) (Thermo, Gibco, MA, USA) and 1% penicillin–streptomycin (Thermo, Gibco, MA, USA). The thyroid cancer cell line FTC133 and normal thyroid cell line NTHY-ORI3-1 were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium containing 10% FBS (Thermo, Gibco, MA, USA) and 1% penicillin–streptomycin (Thermo, Gibco, MA, USA). All cells were cultured at 95% humidity and 5% CO₂ at 37 °C.