Methacholine induced airway contraction in porcine precision cut lung slices from indoor and outdoor reared pigs.

Repetitive exposure to bioaerosols in swine production facilities (SPF) promotes respiratory dysfunction in workers and animals. An adequate understanding of the impact of the SPF environment on pulmonary physiology is needed. However, there is currently no sufficient model to investigate the cause for agriculture-related lung disease. The precision cut lung slices (PCLS) technique represents a practical and useful procedure for studies. Our goal was to use the PCLS technique to develop a model of agriculture-related lung diseases using a physiologically relevant animal model, the domesticated pig. Freshly prepared pig lung tissue cores were sectioned into 300 µm slices and viability was measured by lactate dehydrogenase activity and live/dead staining. Airway contractility in response to a methacholine (MCh) dose gradient (10-10 M) was measured. After the last MCh dose, PCLS were incubated with 1 mM chloroquine to allow airways to relax. Time-lapse images were taken every minute for 35 minutes and used to determine airway lumen area changes. Porcine PCLS remained viable and demonstrated metabolic activity for three days. PCLS from indoor and outdoor pigs contracted in response to MCh exposure and relaxed when incubated with chloroquine. Notably, PCLS of indoor pigs showed greater airway constriction in response to 10 M MCh exposure compared to outdoor pig PCLS (P<0.05). These data suggest that exposure to the indoor pig production environment may be associated with hyperresponsiveness in swine airways, and support future studies to investigate lung response to inflammatory substances using the porcine PCLS model.