Tissue collection and preparation
Regions of lungs were taken from uninvolved areas of neoplastic lesions and resected from 16 patients undergoing lobectomy surgery for lung cancer. Tissues were placed in Krebs-Henseleit buffer solution (KH) as previously described (Cazzola et al., 2011) and transported to the Laboratory of Respiratory Clinical Pharmacology at the University of Rome “Tor Vergata” (Italy) from a nearby hospital. None of the patients had been chronically treated with bronchodilators or corticosteroids, and serum immunoglobulin E (IgE) levels were in the normal range (<100 IU/ml). Preoperative lung function parameters were normal in all the patients who were not affected by chronic obstructive respiratory disorders. Detailed demographic and metric characteristics of donors are reported in e-Table 1.
In the laboratory, the airways were cut into rings (subsegmental bronchi: thickness 1-2 mm, diameter 4-6 mm) and transferred into a 10-ml High Tech 8 Channels Manual Compact Organ Bath system (Panlab Harvard Apparatus, Spain) containing KH buffer solution (37°C) and aerated with O2/CO2 (95:5%). Tissues were allowed to equilibrate and the KH buffer solution was constantly changed.