Msto 211h

Human epithelioid MPM cells, NCI-H226, and human biphasic MPM cells, MSTO-211H, were obtained from American Type Culture Collection (Rockville, MD, United States). RPMI-1640 medium (Gibco, Grand Island, NY) was used for H226 and MSTO-211H cells. The culture media used for the experiments were supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin‒streptomycin (Invitrogen, Carlsbad, CA). Cells were cultivated in the medium at 37 °C in a humidified environment of 5% CO₂.

MPM cell line NCI-H2052 (epithelioid), MSTO-211H (biphasic), NCI-H28 (sarcomatoid), and normal mesothelium cell line Met-5A were purchased from ATCC, US. Met-5A cells were cultured in Medium-199 (Gibco, US) with 10% fetal bovine serum (Gibco, US), and NCI-H2052, MSTO-211H cells NCI-H28 were cultured in RPMI-1640 (Gibco, US) with 10% fetal bovine serum (Gibco, US). Cells were cultured in an incubator under 5% CO2 at 37 °C.

MSTO-211H, NCI-H2452, and 293 T cell lines [American Type Culture Collection (ATCC), Manassas, VA, USA] were employed for this study. MSTO-211H and H2452 were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA), while 293 T cells were cultured in Dulbecco’s modified Eagle’s medium and high glucose medium supplemented with 10% fetal bovine serum. To establish cisplatin-resistant subclones, MSTO-211H and NCI-H2452 cells were cultured with step-wise increasing concentrations of cisplatin [concentrations from the half-maximal inhibitory concentration (IC50) to 10 folds of IC50 of wildtype cells] for 15 months, and 2 cell cisplatin-resistant lines, MSTO- DDP and H2452-DDP, were established. All cells were cultured in a humidified 37 °C incubator with 5% CO₂.