Collection and cultivation of and phagocytosis by pulmonary macrophages obtained from hysterectomy-derived pigs

Abstract

Methods were developed for procuring phagocytically active macrophages from porcine lung with minimal damage to respiratory tissues. Procedures included anesthetizing, surgically introducing a T-shaped tracheal catheter, and repeatedly flushing the respiratory tract. Macrophages collected in this manner were characterized as to numbers, types, and phagocytic activity, nonselective lavage of the pulmonary airways of unstimulated and stimulated (evoking agent: thioglycolate medium) animals yielded 5 X 10(7) and 11 X 10(7) respiratory cells per pig, respectively. Because sufficient quantities (300 to 600 cells/test) of unstimulated cells were collected, stimulated cells contaminated with thioglycolate were not further tested. Morphologically, unstimulated macrophages were mainly spherical and mononucleated by variable in size, ranging from 9 to 30 micrometer. Culturally, macrophages adhered to plastic or glass surfaces and readily phagocytized fungal spores, staphylococci, and latex particles in an enrichment medium containing greater than or equal to 20% bovine fetal serum. Macrophages failed to replicate during a 3-week maintenance period. The data suggest that porcine phagocytes of the pulmonary system comprise a free-cell population that is a major surface-constitutive part of the luminal surface of the airways.