Endothelial cells were isolated from freshly obtained human umbilical cords by collagenase digestion of the interior of the umbilical vein. The cells were grown in tissue culture as a homogeneous population for periods up to 5 mo and some lines were subcultured for 10 serial passages. During the logarithmic phase of cell growth, cell-doubling time was 92 h. Light, phase contrast, and scanning electron microscopy demonstrated that cultured human endothelial cells grew as monolayers of closely opposed, polygonal large cells whereas both cultured human fibroblasts and human smooth muscle cells grew as overlapping layers of parallel arrays of slender, spindle-shaped cells. By transmission electron microscopy, cultured endothelial cells were seen to contain cytoplasmic inclusions (Weibel-Palade bodies) characteristic of in situ endothelial cells. These inclusions were also found in endothelial cells lining umbilical veins but were not seen in smooth muscle cells or fibroblasts in culture or in situ. Cultured endothelial cells contained abundant quantities of smooth muscle actomyosin. Cultured endothelial cells also contained ABH antigens appropriate to the tissue donor's blood type; these antigens were not detectable on cultured smooth muscle cells or fibroblasts. These studies demonstrate that it is possible to culture morphologically and immunologically identifiable human endothelial cells for periods up to 5 mo.

/pdf/culture-of-human-endothelial-cells-derived-from-umbilical-3vt4ihjkgj.pdf

Culture of Human Endothelial Cells Derived from Umbilical Veins. IDENTIFICATION BY MORPHOLOGIC AND IMMUNOLOGIC CRITERIA

CARDIOVASCULAR disease remains the chief cause of death in the United States and Western Europe, and atherosclerosis, the principal cause of myocardial and cerebral infarction, accounts for the majority of these deaths.1 This review, like its predecessor,2 will not attempt to cover all literature on atherosclerosis. In a previous review of the pathogenesis of atherosclerosis,2 Glomset and I discussed various hypotheses of atherogenesis2 3 4 5 6 7 and emphasized the importance of intimal smooth-muscle proliferation as the key event in the development of the advanced lesions of atherosclerosis. The response-to-injury hypothesis of atherogenesis2 3 4 5 6 proposes that "injury" to the endothelium is the initiating event in . . .

The Pathogenesis of Atherosclerosis — An Update

Omega-3 fatty acids in health and disease and in growth and development

We examined whether the synthesis of interleukin-1 or tumor necrosis factor, two cytokines with potent inflammatory activities, is influenced by dietary supplementation with n-3 fatty acids. Nine healthy volunteers added 18 g of fish-oil concentrate per day to their normal Western diet for six weeks. We used a radioimmunoassay to measure interleukin-1 (IL-1 beta and IL-1 alpha) and tumor necrosis factor produced in vitro by stimulated peripheral-blood mononuclear cells. With endotoxin as a stimulus, the synthesis of IL-1 beta was suppressed from 7.4 +/- 0.9 ng per milliliter at base line to 4.2 +/- 0.5 ng per milliliter after six weeks of supplementation (43 percent decrease; P = 0.048). Ten weeks after the end of n-3 supplementation, we observed a further decrease to 2.9 +/- 0.5 ng per milliliter (61 percent decrease; P = 0.005). The production of IL-1 alpha and tumor necrosis factor responded in a similar manner. Twenty weeks after the end of supplementation, the production of IL-1 beta, IL-1 alpha, and tumor necrosis factor had returned to the presupplement level. The decreased production of interleukin-1 and tumor necrosis factor was accompanied by a decreased ratio of arachidonic acid to eicosapentaenoic acid in the membrane phospholipids of mononuclear cells. We conclude that the synthesis of IL-1 beta, IL-1 alpha, and tumor necrosis factor can be suppressed by dietary supplementation with long-chain n-3 fatty acids. The reported antiinflammatory effect of these n-3 fatty acids may be mediated in part by their inhibitory effect on the production of interleukin-1 and tumor necrosis factor.

/pdf/the-effect-of-dietary-supplementation-with-n-3-4mp2ffbrc9.pdf

The effect of dietary supplementation with n-3 polyunsaturated fatty acids on the synthesis of interleukin-1 and tumor necrosis factor by mononuclear cells

Smooth muscle derived from the inner media and intima of immature guinea pig aorta were grown for up to 8 wk in cell culture. The cells maintained the morphology of smooth muscle at all phases of their growth in culture. After growing to confluency, they grew in multiple overlapping layers. By 4 wk in culture, microfibrils (110 A) appeared within the spaces between the layers of cells. Basement membrane-like material also appeared adjacent to the cells. Analysis of the microfibrils showed that they have an amino acid composition similar to that of the microfibrillar protein of the intact elastic fiber. These investigations coupled with the radioautographic observations of the ability of aortic smooth muscle to synthesize and secrete extracellular proteins demonstrate that this cell is a connective tissue synthetic cell.

https://rupress.org/jcb/article-pdf/50/1/172/1385553/172.pdf

The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers.

Several studies have shown that diet-induced non-complicated atherosclerotic lesions regress after removal of the dietary stimulus. Such regression has been reported in monkeys (Armstrong et al., 1970; Armstrong and Megan, 1972; Tucker et al., 1971; Eggen et al., 1974; De Palma et al., 1972; Vesselinovitch et al., 1973), in chickens (Pick et al., 1952; Horlick and Katz, 1949), in pigeons (Clarkson et al., 1971; St. Clair et al., 1972), and in dogs (Bevans et al., 1951; De Palma et al., 1970). In rabbits, regression of atherosclerotic lesions appears to occur only when produced by relatively short-term cholesterol feeding (Bortz, 1968). No regression or only minor shrinkage of lesions resulting from long-term cholesterol feeding has been reported in this animal (Friedman and Byers, 1963; Prior and Ziegler, 1965; McMillan et al., 1955). On the contrary, in some instances, withdrawal of the dietary stimulus led to progression of the disease (Constantinides, 1965; Constantinides et al., 1960). Prior to our study, regression of advanced necrotic lesions (atheromata) and their complications, such as calcification, hemorrhage and thrombosis, has not yet been completely documented. Some studies of advanced lesions suggest that partial regression may occur; however, necrosis and calcification remain (Horlick and Katz, 1949; Prior and Ziegler, 1965).

Regression of Advanced Atherosclerosis in Swine

Atherosclerosis produced in the abdominal aortas of swine by balloon injury and six-month feeding of a high-cholesterol diet resulted in a spectrum of lesions that varied, by gross and/or microscopic examination, from early nonelevated fatty streaks to markedly advanced necrotic and calcified atheromas. On gross examination, the flat lesions had disappeared within five months on the regression regimen, while the advanced lesions persisted after 14 months of withdrawal of the dietary stimulus. Microscopically, the advanced lesions showed changes compatible with a healing process, with a virtual disappearance of foam cells, a marked decrease in necrosis, and replacement of necrotic debris by fibrous tissue and calcification that increased during the regression period. In the early phase of the regression process, the decrease in the number of foam cells was accompanied by an increase in the number of macrophages. The latter cells were found to be closely associated with necrosis, which suggests that macrophages may play a role in the removal of necrotic debris. These results, together with those of our previous experiment on regression of less advanced lesions, suggest that the rate and degree of regression of an atheroma may be a function of its severity.

Sequential morphologic studies of regression of advanced atherosclerosis.

Rapid production of advanced atherosclerosis in swine by a combination of endothelial injury and cholesterol feeding.

Association of DNA synthesis and apparent dedifferentiation of aortic smooth muscle cells in vitro

“Preatheroma” phase of coronary atherosclerosis in man

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