Primary culture of endothelial cells from atherosclerotic human aorta. Part 1. Identification, morphological and ultrastructural characteristics of two endothelial cell subpopulations

Endothelial cells (EC) were harvested by 0.1% collagenase treatment for adult human thoracic aortas obtained 1-3 h after sudden death. At least 35-70% of EC were removed from the intimal surface of aorta, 90-95% of them being viable. Plating efficiency was 70-80%. Monolayer formation was achieved at...

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Bibliographic Details
Published in:	Atherosclerosis Vol. 59; no. 1; p. 1
Main Authors:	Antonov, A S, Nikolaeva, M A, Klueva, T S, Romanov YuA, Babaev, V R, Bystrevskaya, V B, Perov, N A, Repin, V S, Smirnov, V N
Format:	Journal Article
Language:	English
Published:	Ireland 01-01-1986
Subjects:	Adult Aged Aorta, Thoracic - cytology Aorta, Thoracic - pathology Arteriosclerosis - pathology Cell Nucleus Cells, Cultured - classification Child, Preschool Endothelium - cytology Endothelium - immunology Endothelium - pathology Endothelium - ultrastructure Factor VIII - immunology Humans Inclusion Bodies - pathology Infant Male Microscopy, Electron, Scanning Middle Aged Umbilical Veins - cytology
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Summary:	Endothelial cells (EC) were harvested by 0.1% collagenase treatment for adult human thoracic aortas obtained 1-3 h after sudden death. At least 35-70% of EC were removed from the intimal surface of aorta, 90-95% of them being viable. Plating efficiency was 70-80%. Monolayer formation was achieved at a seeding density of 5-8 X 10(2) cells/mm2. The cells were identified as endothelium by the presence of Factor VIII antigen, Weigel-Palade bodies and typically endothelial morphology at confluence. Unlike endothelial cultures derived from human umbilical veins and infant aortas, primary cultures obtained from human adult aortas contain multinuclear EC with Factor VIII antigen and Weibel-Palade bodies. The number of multinuclear EC in cultures isolated from aortas affected by atherosclerosis was 2-fold higher (P less than 0.05) than in cultures obtained from grossly normal aortas taken from donors of the same age. EC with numerous lipid inclusions revealed by oil-red-O staining were present in all the EC primary cultures derived from aortas affected by atherosclerosis. No oil-red-O-positive cells were detected among the EC cultured from infant aorta, aorta of young donors, and umbilical vein. An electron microscopic examination of EC from atherosclerotic aorta in culture and in situ failed to reveal any ultrastructural peculiarities distinguishing multinuclear EC from the mononuclear EC.
ISSN:	0021-9150
DOI:	10.1016/0021-9150(86)90027-4