Isolation and Culture of Human Peritoneal Mesothelial Cells

The Institutional Review Board of Yeungnam University Medical Center (IRB No: 2019-04-047 at 21 December 2019) approved HPMC collection using human specimens. Briefly, gastrectomy was considered for all biopsy-confirmed patients with stomach cancer and was performed using partial or total gastrectomy with total or partial omentectomy. The remnant omentum is typically discarded at our center after surgery. Therefore, prior to surgery, patient informed consent was obtained for use of mesothelial cells from the discarded omentum. HPMCs were isolated as previously described [12 (link),13 (link),14 (link)]. In vitro experiments were performed on cells after one to two passages. HPMCs were cultured in M199 medium supplemented with L-glutamine (20 μM, Gibco, Waltham, MA, USA), penicillin-streptomycin (150 units or 150 μg/mL, Gibco), hydrocortisone (0.4 μg/mL, Sigma-Aldrich, St. Louis, MO, USA), apotransferrin (5 μg/mL, Sigma), insulin (5 μg/mL. Sigma) and 10% fetal bovine serum (FBS) (Gibco). Cell cultures were maintained in 100 mm dishes (Nunc, Rochester, NY, USA) in a 5% CO₂ humidified atmosphere incubator (311 Forma Direct Heat CO₂ incubator, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C.