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Release of prostacyclin, endothelium-derived relaxing factor and endothelin by freshly harvested cells attached to microcarrier beads.

Abstract：

:Cultured endothelial cells have been used in the past as a source of endothelium-derived relaxing factor (EDRF) and of prostacyclin (PGI2). Although cell cultures are essential for observation of prolonged exposure to media or when there is delayed response, they are time consuming and sterile conditions are essential. In the present study, we report that endothelial cells, freshly harvested from bovine aortas, readily attached themselves to cytodex-3 microcarrier beads and released an endothelium-derived relaxing factor (EDRF), prostacyclin (PGI2) and increased the amount of cyclic GMP in vascular smooth muscle. Attachment to microcarrier beads was essential since it increased the surface area and the number of attached cells and permitted collection of cell free filtrates because of the formation of dense networks of cells and beads. As a result superfusion of cells and beads on the filter did not dislodge bound cells which remain on the filter. Conditioned filtrates from freshly harvested endothelial cells attached to microcarrier beads caused marked relaxation of endothelium-deprived bovine pulmonary artery strips. The degree of relaxation depended on the number of cells; maximal relaxation occurred with 50 million cells at ED50 of 14 million. High values of cyclic GMP were found in vascular smooth muscle exposed to conditioned filtrate. The calcium ionophore A23187 further increased the amount of cyclic GMP. Large amounts of PGI2 were released by freshly harvested endothelial cells particularly after stimulation with the calcium ionophore. In contrast, endothelin production by freshly harvested cells attached to microcarrier beads was barely detectable after 30 min incubation and was beyond the limit of detection by bioassay procedures. Freshly harvested endothelial cells attached to microcarrier beads appear to be a useful adjunct to tissue cultures under specific experimental conditions.

journal_name

Mol Cell Biochem

journal_title

Molecular and cellular biochemistry

authors

Kibira S,Dudek R,Narayan KS,Bing RJ

doi

10.1007/BF00239544

subject

Has Abstract

pub_date

1991-11-13 00:00:00

pages

75-84

issue

eissn

0300-8177

issn

1573-4919

journal_volume

108

pub_type

杂志文章

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Release of prostacyclin, endothelium-derived relaxing factor and endothelin by freshly harvested cells attached to microcarrier beads.