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Effects of isolation and culture on prostaglandin synthesis by porcine aortic endothelial and smooth muscle cells
Author(s) -
Ager Ann,
Gordon John L.,
Moncada Salvador,
Pearson Jeremy D.,
Salmon John A.,
Trevethick Michael A.
Publication year - 1982
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041100103
Subject(s) - prostacyclin , prostaglandin , endothelium , radioimmunoassay , prostaglandin e , prostaglandin e2 , medicine , subculture (biology) , endocrinology , tissue culture , vascular smooth muscle , endothelial stem cell , biology , chemistry , biochemistry , smooth muscle , in vitro , microbiology and biotechnology
Freshly isolated neonatal porcine aortic tissue (smooth muscle with or without endothelium present) produced ∼ 30 ng/mg wet tissue of 6‐oxo‐prostaglandin F 1α (the stable hydrolysis product from prostacyclin) and ∼ 15 ng/mg of prostaglandin E 2 , as measured by radioimmunoassay after 24 h incubation in culture medium. Primary cultures of porcine endothelial and smooth muscle cells (isolated by enzymic digestion of aortic tissue) exhibited the same pattern of prostaglandin production, but absolute values were greater than for fresh tissue, particularly in the case of endothelium. Subcultures of endothelium produced smaller amounts of prostaglandins, although the pattern remained similar. In contrast, subcultures of smooth muscle cells produced a greater total amount of prostaglandins than did primary cultures, and the main product was prostaglandin E 2 . Experiments with [ 14 C] prostaglandin H 2 or [ 14 C]arachidonic acid confirmed that aortic tissue, cultured endothelium, and primary cultures of aortic smooth muscle cells synthesized prostacyclin, and demonstrated that subcultured smooth muscle cells enzymically isomerised prostaglandin H 2 H to prostaglandin E 2 . Kinetic studies showed that prostaglandin production by cultured vascular cells was transiently increased by subculture or changing the growth medium, and that production per cell declined with increasing cell density. The change in pattern of prostaglandin production during culture was shown to be due to a rapid decline in the rate of prostacyclin production (which apparently began immediarely after tissue isolation), together with a more gradual rise in prostaglandin E 2 production. These results indicate that the amounts and ratios of prostaglandins produced by vascular endothelial and smooth muscle cells are geatly affected by the conditions used to isolate and culture the cells; vascular cells in vivo may similarly alter their pattern of prostaglandin production in response to local cahnges in their environment.