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Fatty acid composition in native and cultured human endothelial cells

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Summary

Endothelial cells from human umbilical veins were isolated by collagenase treatment. Cells were cultured in the presence of either 20% fetal bovine serum (FBS) or 20% human serum (HS). At confluency, endothelial cell lipids were labeled with tracer concentrations of tritiated arachidonic acid, then extracted and separated into lipid subclasses by thin layer chromatography. The fatty acid composition of each lipid class was determined by glass capillary gas-liquid chromatography analysis and compared to that of cells freshly isolated from the cord (NC cells). The fatty acid compositions differed only in phospholipids. Polyunsaturated fatty acids (PFAs), arachidonic, and linoleic acids were depleted in FBS cell phospholipids and replaced by both stearic and oleic acids. No significant difference could be observed between NC cell and HS cell phospholipids. We conclude that PFAs might be decreased in FBS cells because of the relative paucity of PFAs in FBS as compared to HS. It seems therefore more convenient to cultivate endothelial cells in the presence of HS, especially in respect to their phospholipid content of arachidonic acid, which is the physiological reservoir for prostacyclin synthesis.

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References

  1. Moncada, S.; Gryglewski, R.; Bunting, S.; Vane, J. R. An enzyme isolated from arteries transformed prostaglandin endoperoxides to an unstable substance that inhibits platelet aggregation. Nature 263: 663–665; 1976.

    Article  PubMed  CAS  Google Scholar 

  2. Johnson, R. A.; Morton, D. R.; Kinner, J. H.; Gorman, R. R.; McGuire, J. C.; Sun, F. F. The chemical structure of prostaglandin X (prostacyclin). Prostaglandins 12: 915–928; 1976.

    Article  PubMed  CAS  Google Scholar 

  3. Weksler, B. B.; Marcus, A. J.; Jaffe, E. A. Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells. Proc. Natl. Acad. Sci. USA 74: 3922–3926; 1977.

    Article  PubMed  CAS  Google Scholar 

  4. Marcus, A. J.; Weksler, B. B.; Jaffe, E. A. Enzymatic conversion of prostaglandin endoperoxide H2 and arachidonic acid to prostacyclin by cultured human endothelial cells. J. Biol. Chem. 253: 7238–7141; 1978.

    Google Scholar 

  5. Weksler, B. B.; Ley, C. W.; Jaffe, E. A. Stimulation of endothelial cell prostacyclin production by thrombin, trypsin and the ionophore A 23187. J. Clin. Invest. 62: 923–930; 1978.

    PubMed  CAS  Google Scholar 

  6. Hong, S. L. Effect of bradykinin and thrombin on prostacyclin synthesis in endothelial cells from calf and pig aorta and human umbilical cord vein. Thromb. Res. 18: 787–795; 1980.

    Article  PubMed  CAS  Google Scholar 

  7. Thomas, J. M. F.; Chap, H.; Douste-Blazy, L. Calcium ionophore A 23187 induces arachidonic acid release from phosphatidylcholine in cultured human endothelial cells. Biochem. Biophys. Res. Commun. 103: 819–824; 1981.

    Article  PubMed  CAS  Google Scholar 

  8. Hong, S. L.; Deykin, D. Activation of phospholipases A2 and C in pig aortic endothelial cells synthesizing prostacyclin. J. Biol. Chem. 257: 7151–7154; 1982.

    PubMed  CAS  Google Scholar 

  9. Rastogi, B. K.; Nordoy, A. Lipid composition of cultured human endothelial cells. Thromb. Res. 18: 629–641; 1980.

    Article  PubMed  CAS  Google Scholar 

  10. Spector, A. A.; Hoak, J. C.; Fry, G. L.; Denning, G. M.; Stoll, L. L.; Smith, J. B. Effect of fatty acid modification on prostacyclin production by cultured human endothelial cells. J. Clin. Invest. 65: 1003–1012; 1980.

    PubMed  CAS  Google Scholar 

  11. Macarak, E. J.; De La Llera, M.; Kefalides, N. A.; Howard, B. V. Comparison of the intracellular lipids of cultured vascular endothelial cells and fibroblast. In Vitro 15: 936–940; 1979.

    Article  PubMed  CAS  Google Scholar 

  12. Spector, A. A.; Kaduce, T. L.; Hoak, J. C.; Fry, G. L. Utilization of arachidonic and linoleic acids by cultured human endothelial cells. J. Clin. Invest. 68: 1003–1011; 1981.

    Article  PubMed  CAS  Google Scholar 

  13. Jaffe, E. A.; Nachman, R. L.; Becker, C. G.; Minick, C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52: 2745–2756; 1973.

    PubMed  CAS  Google Scholar 

  14. Lagarde, M.; Burtin, M.; Dechavanne, M.; Sicard, B.; Coiffier, B. Dihomogammalinolenic acid (20∶3ω6) is more anti-aggregatory than eicosapentaenoic (20∶5ω3) in a platelet-endothelial cell mixture. Prostaglandins Med. 4:177–183; 1980.

    Article  PubMed  CAS  Google Scholar 

  15. Morrison, W. R.; Smith, L. M. Preparation of fatty acid methyl esters and dim ethylacetals from lipids with boron fluoride-methanol. J. Lipid. Res. 5: 600–608; 1964.

    PubMed  CAS  Google Scholar 

  16. Schlenk, H.; Gellerman, J. L. Esterification of fatty acids with diazomethane on a small scale. Anal. Chem. 32: 1412–1414; 1960.

    Article  CAS  Google Scholar 

  17. Lagarde, M.; Guichardant, M.; Menashi, S.; Crawford, N. The phospholipid and fatty acid composition of human platelet surface and intracellular membranes isolated by high voltage free flow electrophoresis. J. Biol. Chem. 257: 3100–3104; 1982.

    PubMed  CAS  Google Scholar 

  18. Duck-Chong, C. G. 1979. A rapid sensitive method for determining phospholipid phosphorus involving digestion with magnesium nitrate. Lipids 14: 492–497.

    Article  CAS  Google Scholar 

  19. Carlsson, R. N. K.; Estes, T.; De Groot, J.; Holden, J. T.; Ruoslahti, E. High affinity of α foetoprotein for arachidonic and other fatty acids. Biochem. J. 190: 301–305; 1980.

    PubMed  CAS  Google Scholar 

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This work was supported by a grant from the Délégation Générale à la Recherche Scientifique et Technique, Paris, France (79.7.0091).

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Lagarde, M., Sicard, B., Guichardant, M. et al. Fatty acid composition in native and cultured human endothelial cells. In Vitro 20, 33–37 (1984). https://doi.org/10.1007/BF02633329

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