Summary
We found that fetal bovine serum supplementation of culture medium provided limited quantities of linoleic acid, an essential fatty acid, to cells grown in culture (2.8 ± 0.3% of total fatty acids in 12 lots). Supplementation of the medium with additional linoleic acid resulted in altered phospholipid acyl composition in cells of two established lines, A549, a putative model of the pulmonary Type II epithelial cell, and SIRC, a line derived from rabbit corneal epithelium. In particular, linoleic acid supplementation induced a relative increase in disaturated choline phosphoglycerides of 33 and 36%, respectively, in cells of the two lines. This observation may be relevant to design of media for primary culture of Type II cells, in which disaturated phospholipid synthesis is used as an index of differentiated function (surfactant production). Linoleate supplementation did not alter growth or size (protein content) of cells of either line and caused a slight increase in accumulation of neutral lipid, in the form of cytoplasmic droplets, in A549 cells. Supplementation of cell cultures with equivalent concentrations of the nonessential fatty acids palmitic and oleic acid did not significantly alter the growth, morphologic appearance, or lipid composition of the cells. However, it was demonstrated in cells of one line that palmitic acid supplementation temporarily stimulated synthesis of disaturated choline phosphoglyceride from radiolabeled choline.
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Diglio, C. A.; Kikkawa, Y. The type II epithelial cells of the lung. IV. Adaptation and behavior of isolated type II cells in culture. Lab. Invest. 37: 622–631; 1977.
Smith, F. B.; Kikkawa, Y.; Diglio, C. A.; Dalen, R. C. The type II epithelial cells of the lung. IV. Incorporation of3H-choline and3H-palmitate into lipids of cultured type II cells. Lab. Invest. 42: 296–301; 1980.
Gould, K. G., Jr. Dispersal of lung into individual viable cells. Lenfant, C. ed. Lung biology in health and disease. New York: Marcel Dekker; 1976: 49–72.
Kikkawa, Y.; Yoneda, K. The type II epithelial cell of the lung. I. Method of isolation. Lab. Invest. 30: 76–84; 1974.
Kikkawa, Y.; Yoneda, K.; Smith, F.; Packard, B.; Suzuki, K. The type II epithelial cells of the lung. II. Chemical composition and phospholipid synthesis. Lab. Invest. 32: 295–302; 1975.
Mason, R. J.; Williams, M. C.; Greenleaf, R. D.; Clements, J. A. Isolation and properties of type II alveolar cells from rat lung. Am. Rev. Respir. Dis. 115: 1015–1026; 1977.
Smith, B. T. Cell line A549: A model system for the study of alveolar type II cell function. Am. Rev. Respir. Dis. 115: 285–293; 1977.
Mason, R. J.; Williams, M. C. Phospholipid composition and ultrastructure of A549 cells and other cultured pulmonary epithelial cells of presumed type II cell origin. Biochim. Biophys. Acta 617: 36–50; 1980.
Schneeberger, E. E.; Lynch, R. D.; Geyer, R. P. Formation and disappearance of triglyceride droplets in strain L fibroblasts. Exp. Cell. Res. 69: 193–206; 1971.
Spector, A. A. Fatty acid, glyceride, and phospholipid metabolism. Rothblat, G. H.; Cristofalo, V. J. eds. Growth, nutrition, and metabolism of cells in culture. Vol. 1. New York: Academic Press; 1972: 257–296.
Miller, J. S.; Garnio, V. C.; Ackerman, G. A.; Sharma, H. M.; Milo, G. E.; Geer, J. C.; Cornwall, D. G. Triglycerides, lipid droplets and lysosomes in aorta smooth muscle cells during control of cell proliferation with polyunsaturated fatty acids and vitamin E. Lab. Invest. 42: 495–506; 1980.
Holley, R. W.; Baldwin, J. H.; Kiernan, J. A. Control of growth of a tumor cell by linoleic acid. Proc. Natl. Acad. Sci. USA 71: 3976–3978; 1974.
Horwitz, A. F.; Hatten, M. E.; Binger, M. M. Membrane fatty acid replacements and their effect on growth and lectin-induced agglutinibility. Proc. Natl. Acad. Sci. USA 71: 3115–3119; 1974.
Huttner, J. J. Fatty acids and their derivatives: inhibitors of proliferation in aortic smooth muscle cells. Science 197: 289–291; 1977.
Limanek, J. S.; Chin, J.; Chang, T. Y. Mammalian cell mutant requiring cholesterol and unsaturated fatty acid for growth. Proc. Natl. Acad. Sci. USA 75: 5452–5456; 1978.
Spector, A. A.; Kiser, R. E.; Denning, G. M.; Koh, S. M.; DeBault, L. E. Modification of the fatty acid composition of cultured human fibroblasts. J. Lipid Res. 20: 536–547; 1979.
Wicha, M. S.; Liotta, L. A.; Kidwell, W. R. Effects of free fatty acids on the growth of normal and neoplastic rat mammary epithelial cells. Cancer Res. 39: 426–435; 1979.
Lynch, R. D. Utilization of polyunsaturated fatty acids by human diploid cells ageing in vitro. Lipids 15: 412–420; 1980.
Gerschenson, L. E.; Mead, J. F.; Harary, I.; Haggerty, D. F. Studies on the effects of essential fatty acids on growth rate, fatty acid composition, oxidative phosphorylation and respiratory control of HeLa cells in culture. Biochim. Biophys. Acta 131: 42–49; 1967.
Wood, R.; Falch, J. Lipids of cultured hepatoma cells. II. Effect of media lipids on cellular phospholipids. Lipids 8: 702–710; 1973.
MacKenzie, C. G.; Moritz, E.; Wisneski, J. A.; Reiss, O. K.; Moe, K. J. B. Fatty acid ester turnover: a control factor in triglyceride and lipid rich particle accumulation in cultured mammalian cells. Mol. Cell. Biochem. 19: 7–15; 1978.
Lieber, M.; Smith, B.; Szakal, A.; Nelson-Rees, W.; Todaro, G. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer 17: 62–70; 1976.
Leehoy, J. Cytopathic effect ofRubella virus in a rabbit-cornea cell line. Science 149: 633–634; 1965.
Smith, F. B.; Kikkawa, Y. The type II epithelial cells of the lung. III. Lecithin synthesis: a comparison with pulmonary macrophages. Lab. Invest. 38: 45–51; 1978.
Lowry, O. H.; Rosenbrough, N. J.; Farr, A. L.; Randall, R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265–275; 1951.
Bligh, E. G.; Dyer, W. J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37: 911–917; 1959.
Mason, R. J.; Huber, G.; Vaughan, M. Synthesis of dipalmitoyl lecithin by alveolar macrophages. J. Clin. Invest. 51: 68–73; 1972.
Mangold, H. K. Thin layer chromatography of lipids. J. Am. Oil Chem. Soc. 38: 708–727; 1961.
Rouser, G.; Siakotos, A. N.; Fleisher, S. Quantitative analysis of phospholipids and phosphorus analysis of spots. Lipids 1: 85–86; 1966.
Ways, P.; Reed, C. F.; Hanahan, D. J. Red cell and plasma lipids in acanthocytosis. J. Clin. Invest. 42: 1248–1260; 1963.
Lindgren, F. T.; Nichols, A. V. Fatty acid composition of the serum lipoproteins. Ann. N.Y. Acad. Sci. 94: 55–70; 1961.
Springer, E. L. Comparative study of the cytoplasmic organelles of epithelial cell lines derived from human carcinomas and malignant tissues. Cancer Res. 40: 803–817; 1980.
Wilson, J. W.; Leduc, E. H. Mitochondrial changes in the liver of essential fatty acid deficient mice. J. Cell Biol. 16:281–296; 1963.
Gordon, G. B. Saturated free fatty acid toxicity. II. Lipid accumulation, ultrastructural alternations and toxicity in mammalian cells in culture. Exp. Mol. Pathol. 27: 262–276; 1977.
Holman, R. T. Essential fatty acids in human nutrition. Adv. Exp. Med. Biol. 83: 515–534; 1977.
Mathers, L. Enzyme deletions and essential fatty acid metabolism in cultured cells. J. Biol. Chem. 250: 1152–1153; 1975.
Van Golde, L. M. G. Metabolism of phospholipids in the lung. Am. Rev. Resp. Dis. 114: 977–1000; 1976.
Thompson, G. A., Jr. The regulation of membrane lipid metabolism. Boca Raton: CRC Press, Inc.; 1980.
Gordon, G. B. Lipid accumulation in the stationary phase of strain L cells in suspension culture. Lab. Invest. 36: 114–121; 1977.
Gordon, G. B.; Barcza, M. A.; Bush, M. E. Lipid accumulation in hypoxic tissue culture cells. Am. J. Pathol. 88: 663–674; 1977.
Kyriakides, E. C.; Beeler, D. A.; Edmonds, R. H.; Balint, J. A. Alterations in phosphatidyl choline species and their reversal in pulmonary surfactant during essential fatty acid deficiency. Biochim. Biophys. Acta 431: 399–407; 1976.
Burnell, J. M.; Kyriakides, E. C.; Edmonds, R. H.; Balint, J. A. The relationship of fatty acid composition and surface activity of lung extracts. Respir. Physiol. 32: 195–206; 1978.
Puck, T. T.; Cieciura, S. J.; Robertson, A. Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects. J. Exp. Med. 108: 945–956; 1958.
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This work was supported by Grants HL-24817 and HL-21251 from the National Institutes of Health, USPHS, and by a grant from the Alexandrine and Alexander L. Sinsheimer Fund.
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Smith, F.B., Kikkawa, Y., Diglio, C.A. et al. Increased saturated phospholipid in cultured cells grown with linoleic acid. In Vitro 18, 331–338 (1982). https://doi.org/10.1007/BF02796331
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DOI: https://doi.org/10.1007/BF02796331