Design of a new stretching apparatus and the effects of cyclic strain and substratum on mouse lung epithelial-12 cells

The pulmonary epithelium is exposed to mechanical strains during normal breathing or mechanical ventilation. While important for the regulation of cellular processes, excessive strains damage epithelial cells. To investigate the effects of strain on the epithelium, we developed a stretching device to apply equi-biaxial strains to cells cultured on elastic membranes. Following device validation, we exposed a murine epithelial cell line (MLE-12) to 30 min of cyclic stretch with 0, 25, 50, 75 and 100% change in surface area on pronectin or type I collagen coated membranes. Following stretch, we assessed cell viability using fluorescent immunocytochemisty and surfactant secretion using [H-3] labeled phosphatidylcholine (PC). Cell injury increased with increasing strain with cells on pronectin showing more injury than on type I collagen. Stretching had no effect on surfactant secretion on either substratum suggesting MLE-12 cells are a poor model for stretch-induced surfactant secretion. The cells grown on pronectin, however, demonstrated a 3-fold increase in surfactant secretion compared to those grown on type I collagen at all strains. This suggests that, while this cell line does not demonstrate stretch-induced surfactant secretion, the underlying extracellular matrix plays a crucial factor in both cell death and signal transduction in response to strain.