Culturing Peritoneal Tissue Explants

Fresh peritoneal tissue was obtained directly from the operating room, with informed consent from the patient, and transferred immediately to the laboratory in E199 medium (Biochrom AG, Berlin, Germany). The tissue was then incubated for 15 min in phosphate-buffered saline (PBS)-containing penicillin/streptomycin and amphotericin (Sigma–Aldrich Chemie, St. Louis, MO, USA). Extraperitoneal fat was removed, and 7 × 7 mm tissue pieces were inserted between two stainless steel rings and cultured, with the mesothelial cell surface directed upwards, in E199 medium containing penicillin/streptomycin, L-glutamine (Biochrom AG, Berlin, Germany), FCS (Thermo Fisher Scientific GmbH, Waltham, MA, USA), hydrocortisone (Sigma–Aldrich Chemie, St. Louis, MO, USA), fibroblast growth factor (PeproTech GmbH, Hamburg, Germany), and heparin (Biochrom AG, Berlin, Germany), as described previously [12 (link)].