title: Isolation and culture of endothelial cells from the lungs of small animals.
abstract: Techniques are descirbed for the isolation and culture of endothelial cells from the lungs of small animals. The cells are collected by retrograde perfusion of blood-free lungs with buffered saline containing collagenase. The cells are characterized by light microscopy, electron microscopy 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. Thus, endothelial cultures can be established using small animals commonly employed in studies of pulmonary processing of vasoactive substances.
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002875:Chromosomes; D003584:Cytological Techniques; D004727:Endothelium / Q000166:cytology / Q000201:enzymology; D005260:Female; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D008168:Lung / Q000166:cytology* / Q000201:enzymology; D008297:Male; D003012:Microbial Collagenase / Q000494:pharmacology; D008854:Microscopy, Electron; D007703:Peptidyl-Dipeptidase A / Q000032:analysis; D010477:Perfusion / Q000379:methods; D011817:Rabbits; D051381:Rats
chemical_list: D007703:Peptidyl-Dipeptidase A; D003012:Microbial Collagenase