Met5a

The primary pleural mesothelial cells NP2 and its hTERT-transduced derivative NP2-hT⁺ were cultured in ACL growth medium supplemented with 10% FBS. The primary Meso-CAFs (Meso109F, Meso125F) and their hTERT-transduced derivatives (Meso109F-hT⁺, Meso125F-hT⁺) as well as the human mesothelioma cell lines (MSTO-211H, SPC212) were all kept in RPMI-1640 medium with 10% FBS. MSTO-211H was purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA) and SPC212 was kindly provided by Prof. R. Stahel (University of Zurich, Zurich, Switzerland). The SV40-immortalized, non-malignant human pleural mesothelial cell line Met5A and both normal primary human lung fibroblasts, MRC-5 and Wi38, were also obtained from ATCC (Rockville, MD, USA) and cultivated in the respective growth medium according to the supplier’s protocol (Met5A in RPMI-1640, MRC-5 in Dulbecco’s Modified Eagle’s Medium (DMEM) (#D5648, Sigma-Aldrich, St. Louis, MO, USA), and Wi38 in Eagle’s minimal essential medium (MEM) (#M5650, Sigma-Aldrich, St. Louis, MO, USA), all supplemented with 10% FBS). The primary pleural mesothelial cells and the primary Meso-CAFs used for experiments were at fewer than 15 passages. All used cells were maintained in a humidified atmosphere (37 °C, 5% CO₂), regularly passaged by trypsinization, and routinely checked for Mycoplasma contamination.

AB22 murine mesothelioma and MeT-5A human healthy mesothelial cell lines were purchased from Sigma and ATCC respectively. AB22 cells were cultured in RPMI media supplemented with 25 mM Hepes, 10% FBS, 1% P/S and 2 mM of Glutamine, while MeT-5A were cultured in Medium 199 supplemented with 10% FBS, 1% P/S, 8.7E⁻⁴ mM of bovine insulin, 3.3E⁻⁶ mM of human EGF, 4E⁻⁴ mM of hydrocortisone and Trace Elements B (Corning) and maintained in a 5% CO₂ incubator at 37 °C.