Cultivation of HeLa cells with fetal bovine serum or Ultroser G: effects on the plasma membrane constitution.
Plasma membranes isolated from HeLa cells cultivated in suspension cultures supplemented with 3.5% fetal bovine serum or 2% of the commercially available serum substitute Ultroser G contained the same amounts of protein, cholesterol, and phosphate on a cellular basis. Minor differences in the plasma membrane fatty acid composition were seen, with the most pronounced alteration observed for palmitic acid, which amounted to 27 and 20% in fetal bovine serum- and Ultroser G- supplemented cells, respectively. Plasma membranes from cells grown with Ultroser G contained almost twice as much phosphatidylethanolamine and displayed two thirds of the phosphatidylcholine content, compared to plasma membranes obtained from fetal bovine serum supplemented cells. The former membranes also showed a 3 times higher specific [3H]acetate labeling of cholesterol, indicating a higher de novo synthesis of cholesterol. Both quantitative and qualitative alterations were revealed among the plasma membrane polypeptides when these were subjected to immuno- and lectin blottings. Fluorescence anisotropy measurements at different temperatures produced similar results irrespective of the growth medium supplement when the plasma membrane specific probe 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene was used on intact cells. However, the average cellular rigidity was higher for Ultroser G supplemented cells, determined with 1,6-diphenyl-1,3,5-hexatriene as a probe.
Blixt
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Y
Department of Microbiology, University of Lund, Sweden.
Valeur
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A
Everitt
E
E
D001769:Blood; D001802:Blood Substitutes / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D005227:Fatty Acids / Q000378:metabolism; D005454:Fluorescence Polarization; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D006801:Humans; D015151:Immunoblotting; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009930:Organic Chemicals; D010710:Phosphates / Q000378:metabolism
D001802:Blood Substitutes; D003470:Culture Media; D005227:Fatty Acids; D008563:Membrane Lipids; D008565:Membrane Proteins; D009930:Organic Chemicals; D010710:Phosphates; C044609:Ultroser G; D002784:Cholesterol