title: Development and characterization of essential fatty acid deficiency in human endothelial cells in culture.
abstract: We induced an essential fatty acid deficiency (EFAD) in human umbilical vein endothelial cells by culture in medium with 20% (vol/vol) delipidated fetal calf serum. EFAD, reflected by decreased cellular linoleic acid (18:2 omega 6) and arachidonic acid (20:4 omega 6) and emergence of the oleic acid derivative 5,8,11-eicosatrienoic acid (20:3 omega 9; Mead's acid), was evident after 1 week of culture and became pronounced after 2 weeks. Beyond that time point, control cells (cultured in 20% normal fetal calf serum) grew deficient of 18:2 omega 6, and EFAD cells died. 18:2 omega 6 addition to EFAD cells resulted in dose-dependent increases of 18:2 omega 6 and 20:4 omega 6. 20:4 omega 6 or 5,8,11,14,17-eicosapentaenoic acid (20:5 omega 3) additions resulted in normalization of these acids, and conversion of 20:5 omega 3 to 4,7,10,13,16,19-docosahexaenoic acid (22:6 omega 3) was noted. Agonist-induced increases in concentrations of prostacycline (prostaglandin I2; PGI2) and cytosolic Ca2+, [Ca2+]i, were reduced in EFAD cells and not restored by 18:2 omega 6 or 20:4 omega 6 additions. Change of the medium in EFAD cultures 1 day before the experiments decreased 20:3 omega 9 and normalized the PGI2 production and [Ca2+]i changes, whereas addition of 20:3 omega 9 to control cells impaired the [Ca2+]i response, indicating a suppressive effect of 20:3 omega 9. Thus, EFAD in endothelial cells is associated with abnormalities of eicosanoid and second-messenger production partly attributable to 20:3 omega 9 accumulation. Moreover, the gradual emergence of 18:2 omega 6 deficiency in regularly grown control cells underlines the need for careful analysis of fatty acids in long-term cell cultures.
authors.affiliation: Department of Medicine, Karolinska Institute, Stockholm Söder Hospital, Sweden.
authors.forename: R
authors.initials: R
authors.lastname: Lerner
authors.forename: P
authors.initials: P
authors.lastname: Lindström
authors.forename: A
authors.initials: A
authors.lastname: Berg
authors.forename: E
authors.initials: E
authors.lastname: Johansson
authors.forename: K
authors.initials: K
authors.lastname: Rosendahl
authors.forename: J
authors.initials: J
authors.lastname: Palmblad
mesh_terms: D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D011464:Epoprostenol / Q000378:metabolism; D005228:Fatty Acids, Essential / Q000172:deficiency* / Q000378:metabolism; D006801:Humans; D008052:Lipid Metabolism, Inborn Errors / Q000378:metabolism*
chemical_list: D005228:Fatty Acids, Essential; D011464:Epoprostenol; D002118:Calcium