A Platelet-Dependent Serum Factor That Stimulates the Proliferation of Arterial Smooth Muscle Cells In Vitro

Abstract

Dialyzed serum from clotted monkey blood ("blood serum") promotes the proliferation of monkey arterial smooth muscle cells in culture, but dialyzed serum prepared from recalcified platelet-poor plasma ("plasma serum") is much less effective. Addition of platelets and calcium to platelet-poor plasma increases the activity of plasma serum to the same level achieved with blood serum. Furthermore, addition to plasma serum of a platelet-free supernatant prepared by exposing purified platelets to thrombin also stimulates the proliferation of smooth muscle cells. Thus, much of the growth-promoting activity of dialyzed serum is directly or indirectly derived from platelets. This finding has important implications for the response of arteries to localized injury and provides a key to further understanding of the role of factors derived from blood serum in promoting cell proliferation in vitro.We have been studying the growth of arterial smooth muscle cells (SMC) in culture as part of an attempt to determine why these cells accumulate focally in atherosclerosis (1). Since our working hypothesis has been that atherosclerosis is an exacerbated arterial response to local endothelial injury, we have been interested in the fact that SMC, like most other diploid, nontransformed cells, do not proliferate in culture except in the presence of blood serum. We have been attempting to identify the serum factors involved, in order ultimately to test the possibility that endothelial injury increases their concentration in the subendothelial space and thereby promotes SMC proliferation in the intima. We have already found (1, 2) that serum lipoproteins are necessary for optimal cell growth in culture, and now wish to report studies of a nondialyzable serum component that is probably not a lipoprotein and that appears to be derived from platelets. MATERIALS AND METHODS Procedures described in detail elsewhere (1-3), were used to subculture SMC from explants of the intima and inner media of the thoracic aorta of pigtail monkeys (Macaca nemestrina). The explants were grown in a modified Dulbecco-Vogt medium (3) based on that originally described by Eagle, and containing 5% or 10% pooled blood serum from the same species of monkey. SMC from the explants were subcultured in a similar medium that contained 5% blood serum and lacked streptomycin. Cells from the third to sixth subculture were used to test the effect of serum factors upon cell proliferation. The assay procedure used was similar to that of Paul et al. (4) equal numbers in 35-mm Falcon plastic petri dishes. The reproducibility of cell plating was confirmed in each experiment. The medium was changed three times per week. For the first 7 days the medium .contained 1% 'pooled monkey blood serum. Thereafter, the 1% serum was replaced with the different serum fractions to be tested. Each fraction was added in an amount that corresponded to that provided by 5% monkey blood serum, and subsequent cell proliferation was compared with that obtained in medium containing 5% blood ser...

Animal‐cell culture media: History, characteristics, and current issues

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Nox5 mediates PDGF-induced proliferation in human aortic smooth muscle cells

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Platelet-derived growth factor enhances in vitro erythropoiesis via stimulation of mesenchymal cells.

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