Dialyzable factor in human serum of platelet origin stimulates endothelial cell replication and growth |
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| Authors: | David R Clemmons William L Isley and M Todd Brown |
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| Affiliation: | 1Department of Medicine, Center for Thrombosis and Hemostasis, University of North Carolina, Chapel Hill, North Carolina 27514 |
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| Abstract: | Porcine aortic endothelial cells were isolated and maintained in Dulbecco's modified Eagle's medium (DME medium)/10% citrate-treated human plasma. They were stimulated by DME medium/10% human serum to grow from a density of 10,100 ± 500 per well to a final density of 83,000 ± 1,800 per well over a 9-day period. On the other hand these cells grew poorly (11% increase) in DME medium/10% human platelet-poor plasma prepared without chelating agents and containing platelet factor 4 at 18 ng/ml by radioimmunoassay. Dialysis of the human serum (Mr cutoff, 3,500) eliminated all the stimulatory activity. The activity recovered from the dialysate stimulated growth when added to endothelial cultures in conjunction with either dialyzed serum or platelet-poor plasma alone. The dialyzable factor could be obtained directly from platelets; both acetic acid extracts and boiled NaCl extracts stimulated porcine aortic endothelial cell replication. Gel filtration chromatography on Sephadex G-15 showed that the endothelial growth factor had a molecular weight of 700. Partially purified material induced a concentration-dependent stimulation of porcine aortic endothelial cell replication in the presence of DME medium alone; however, simultaneous incubation with platelet-poor plasma resulted in a much greater response. Fibroblast growth factor isolated from bovine brain was found to be mitogenic only in the presence of nondialyzed serum or of the dialyzable factor together with plasma. In the absence of this serum factor, fibroblast growth factor had no effect. We conclude that human serum contains a potent endothelial cell mitogen of platelet origin. Human plasma that is devoid of platelet content does not stimulate endothelial cell growth. This growth factor may be an important stimulant of the endothelial cell response to vascular wall injury. |
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