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

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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.

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