Porcine airway tissue explant culture at a hydrogel-air interface to model submucosal glands in cystic fibrosis progression and therapy

Submucosal glands (SMGs) are critical for airway health; however, replicating their complex structure and function outside the body remains challenging. Here, we establish a method of airway tissue explant culture at a hydrogel-air interface (i.e., AirTECH) using trachea from newborn pigs. After 14 days, AirTECH tissues maintain their natural SMG structures, cell types, gene expression, and defense functions, along with intact surface epithelia, smooth muscle, and cartilage. Using AirTECH tissue as a model, we evaluated SMGs in cystic fibrosis (CF) airway disease progression, where exposure to lipopolysaccharide of Pseudomonas aeruginosa caused abnormal mucus obstruction and overexpression of MUC5AC in SMGs, a characteristic feature of CF airway disease. We also explored the contribution of SMGs in airway gene therapy, where green fluorescent protein–encoding adenovirus vectors were transduced and expressed exclusively in airway surface epithelia but not SMGs. These data support AirTECH as a model to understand SMGs in airway disease and test therapeutic interventions.