ISOLATION AND CULTURE OF ENDOTHELIAL-CELLS FROM THE LUNGS OF SMALL ANIMALS

Abstract

Techniques for the isolation and culture of endothelial cells from the lungs of small animals were described. The cells were collected by retrograde perfusion of blood-free lungs with buffered saline containing collagenase, and were characterized by light microscopy, EM of thin sections and surface replicas and by the presence of angiotensin-converting enzyme (ACE). ACE was assayed using 3H-benzoyl-Phe-Ala-Pro as substrate and was localized by indirect immunofluorescence using guinea pig endothelial cells incubated with rabbit antibodies to guinea pig lung ACE followed by goat anti-rabbit globulins conjugated to fluorescein. Endothelial cultures were established using small animals commonly employed in studies of pulmonary processing of vasoactive substances.