MPM cells were isolated from pleural biopsies of ten enrolled patients following the previously reported procedure [20 (link)]. MPM primary cells were cultured in a Human Endothelial Serum Free Medium (HESFM, Gibco) supplemented with 20 ng/mL epidermal growth factor (Sigma), 10 ng/mL basic fibroblast growth factor (Immunological Sciences), 1% penicillin–streptomycin (Sigma-Aldrich) and 10% heat-inactivated fetal bovine serum (FBS, Gibco). The cells were maintained at 37 °C in humidified atmosphere with 5% v/v CO2 and the medium was changed every 2–3 days. To assess the purity of isolated primary cells, they were characterized both by cytofluorimetric analysis and immunofluorescence assays for the following MPM representative markers: mesothelin, calretinin, cytokeratin 8/18, WT1 and CD44 in addition to CD45 and vWF to exclude leukocyte and endothelial cell contamination, respectively, as previously described [20 (link)].
Met5A cells, a non-malignant human pleural mesothelial line, were purchased from the American Type Culture Collection (ATCC) and cultured in complete HESFM. NCI-H28 cells, a human mesothelioma cell line, were purchased from the ATCC and cultured in the RPMI-1640 medium supplemented with 10% FBS.
Met5A cells, a non-malignant human pleural mesothelial line, were purchased from the American Type Culture Collection (ATCC) and cultured in complete HESFM. NCI-H28 cells, a human mesothelioma cell line, were purchased from the ATCC and cultured in the RPMI-1640 medium supplemented with 10% FBS.