Ultrastructural and immunohistochemical studies of primary cultures of aortic medial cells

Fisher-Dzoga, Katti; Jones, Rose M.; Vesselinovitch, Dragoslava; Wissler, Robert W.

doi:10.1016/0014-4800(73)90014-2

Cited by 133 publications

(30 citation statements)

References 17 publications

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“…Cells were identified as smooth muscle on the basis of growth characteristics in culture and on the basis of ultrastructural examination. Methods for cell preparation, criteria for identification, and description and quantitation of smooth muscle cell features have been presented elsewhere for cells grown from explants in our tissue culture laboratories 21 and for cells harvested from elastin membranes in our cell stretching studies.…”

Section: ^0mentioning

confidence: 99%

Effect of normolipemic and hyperlipemic serum on biosynthetic response to cyclic stretching of aortic smooth muscle cells.

et al. 1989

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Arterial smooth muscle cells synthesize matrix macromolecules In response to mechanical stimulation. Exposure to serum llpids also stimulates connective tissue fiber accumulation. To assess the effect of serum llplds on the blosynthetic response to tensile stress, we subjected rabbit aortic smooth muscle cells that were cultured on purified elastin membranes to cyclic stretching and relaxation 50 times per minute In the presence of serum-free medium (SFM), normolipemic serum (NLS), or hyperlipemic serum (HLS). Incorporation of 14 C-prollne Into prollne and Into hydroxyprollne was taken as a measure of protein and collagen synthesis. When cells were grown in plastic Petrl dishes, exposure to NLS or HLS Increased both protein and collagen production to the same extent compared to synthesis In SFM (1.7 times for NLS and 1.6 times for HLS; p<0.001 compared to SFM). For cells grown on stationary elastin membranes, NLS and HLS also Increased protein and collagen synthesis compared to SFM. The effect of NLS was 1.35 times that of HLS for protein and 1.43 times greater for collagen (p<0.03). Cyclic stretching In SFM doubled synthesis for both protein (p<0.002) and collagen (p<0.002) compared to stationary controls, but had no effect on synthesis In NLS. In HLS, however, cyclic stretching elevated synthesis to the same level as was found In NLS (p<0.003). We conclude that the relative Inhibition of synthesis on stationary membranes by HLS was not due to a toxic effect, since HLS Increased synthesis both In Petrl dishes and on elastin membranes, and the amplifying effect of cyclic stretching In HLS was similar to that seen In SFM. The Interactive effects of mechanical stimulation and hyperllpemla may help to account for Individual differences In wall and plaque composition during atherogenesls. (Arteriosclerosis 9:446-452, July/August 1989) S mooth muscle cells may modify their macromolecular microenvironment by synthesizing elastin, collagen, and glycosaminoglycans. 1 • 23 Some of these changes have been associated with mechanical stimuli. During early growth, for example, differences in the rate of matrix fiber accumulation between the ascending aorta and the pulmonary trunk correspond closely to the respective rates of increase of mural tensile stress despite the persistent similarity of cell content for the two vessel segments. 4 In adult mammals, the number and microarchitecture of the musculoelastic layers of the media of homologous arteries is related to vessel diameter and curvature 56 and, therefore, to the distribution of tensile stress in the vessel wall. Under conditions of hypertension, the number of transmural layers may not change, but the cross-sectional area of the media increases in propor-

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Section: ^0mentioning

confidence: 99%

Effect of normolipemic and hyperlipemic serum on biosynthetic response to cyclic stretching of aortic smooth muscle cells.

et al. 1989

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“…Arterial medial smooth muscle cells were obtained from outgrowths of explants from the thoracic aorta of Macaca mulatta monkeys (rhesus) as previously described, 22 -23 grown in Basal Medium of Eagle (BME) with 5% calf serum, and used between the 4th and 6th passages (0.3-1.0 mg protein/flask). Experiments were carried out in 25 cm 2 flask (Falcon) in a volume of 2.0 ml per flask and terminated as described for macrophages.…”

Section: Tissue Culturementioning

confidence: 99%

Very low density lipoproteins promote triglyceride accumulation in macrophages.

Bates¹,

Murphy²,

Zhou³

et al. 1984

Arteriosclerosis

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Incubation of mouse peritoneal macrophages with very low density lipoproteins (VLDL) from normal rats or rhesus monkeys markedly increased the levels of intracellular trlglycerldes by 10-to 56-fold and was accompanied by the production of oil red 0 positive vacuoles. The stimulation of triglyceride accumulation in macrophages was time-and concentration-dependent and was specific for VLDL. Three possible mechanisms for the VLDL-stlmulated triglyceride accumulation In macrophages were explored: receptor-mediated uptake, action of llpoprotein llpase, and phagocytosis. Macrophage uptake and degradation of 125 l-monkey and rat VLDL demonstrated saturable and nonsaturable components. Uptake of 125 I-VLDL could be inhibited by unlabeled normal VLDL, although hyperllpemlc VLDL was more effective. HDL did not compete to a significant extent. Heparln released llpoprotein lipase-llke activity from peritoneal macrophages. Addition of heparln with VLDL resulted in a greater, more rapid elevation in Intracellular triglycerides, which was partially Inhibited by albumin. Free fatty acid and Intrallpid also produced triglyceride accumulation In macrophages. The data showed that all three of the mechanisms examined could contribute to the metabolism of VLDL by macrophages and cause the production of trlglycerlderlch cells with a "foamy" appearance, although the evidence suggested that the action of llpoprotein llpase was probably the most important In this process.

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“…11 These proliferating SMC synthesize extracellular matrix materials, such as collagen 12 and glycosaminoglycans; 13 their growth is stimulated by several factors such as those found in platelets 14 ' 15 and in hyperlipemic, 16 diabetic, 17 and hypertensive 18 sera. Our working hypothesis is that the readiness of SMC to migrate onto a culture dish and to proliferate in vitro is a measurement of the extent of pathologi-.…”

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confidence: 99%

Intimal injury in vivo activates vascular smooth muscle cell migration and explant outgrowth in vitro.

Grünwald¹,

Haudenschild²

1984

Arteriosclerosis

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In vivo induced endothelial injury results in an intimal thickening, mainly caused by stimulation of smooth muscle cells (SMC), which migrate into, and proliferate in, the denuded intima. We evaluated the manifestation and persistence of this stimulus in several in vitro explant and growth assays using aortic explants from either balloon catheter-injured rats or from sham-operated controls. SMC outgrowth from control explants started 48 hours after explantation, became half-maximal after 96 hours, with 74% of the explants showing outgrowth. Explants from balloon-injured aortas showed SMC outgrowth within 24 hours, became half-maximal after 48 hours, with 92% of the explants showing outgrowth. An interval of 4 to 7 days between balloon injury and sacrifice was optimal for obtaining accelerated outgrowth, while a 1-day interval showed a weak stimulation and a 21-day interval had no such effect. The in vivo stimulated cells grew more rapidly, resulting in a greater colony size per explant, and they became temporarily serum-independent. Increased growth rate persisted up to the second subculture but returned to normal in subsequent passages. 2 Altered endothelial integrity is thought to expose the underlying SMC to stimuli that induce migration of the SMC through the internal elastic lamina to the intima, where they proliferate.34 Approximately 3 to 4 days after experimental injury, 56 activated or modulated SMC 7 ' 8 appear at the luminal surface and form an intimal thickening that displays many features of an atherosclerotic plaque.Cell culture techniques are now widely used to study the growth kinetics and morphology of SMC. 24 Differences in SMC outgrowth after mechanical injury of the rabbit aorta have also been reported briefly. 25 We have extended this work in the studies reported here by quantifying SMC migration and proliferation in explant cultures and subcultivations after balloon injury of the rat aorta. These studies show that SMC migration and proliferation in vitro are accelerated and enhanced in response to the in vivo injury. Furthermore, the in vivo stimulus renders the SMC temporarily serum-independent.

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Ultrastructural and immunohistochemical studies of primary cultures of aortic medial cells

Cited by 133 publications

References 17 publications

Effect of normolipemic and hyperlipemic serum on biosynthetic response to cyclic stretching of aortic smooth muscle cells.

Effect of normolipemic and hyperlipemic serum on biosynthetic response to cyclic stretching of aortic smooth muscle cells.

Very low density lipoproteins promote triglyceride accumulation in macrophages.

Intimal injury in vivo activates vascular smooth muscle cell migration and explant outgrowth in vitro.

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