Keeping the intracellular vitamin C at a physiologically relevant level in endothelial cell culture.
It is generally accepted that the addition of vitamin C to cell culture medium improves cell growth. However, once added, the vitamin C concentration declines rapidly. This situation differs from the in vivo environment where the endothelium is constantly supplied with ascorbate from the blood. With a focus on intracellular vitamin C, we simulated constant supply of ascorbate by the hourly addition of freshly prepared medium containing 75 microM ascorbate and subsequently compared it with more practical regimens using combinations of ascorbate and 2-phosphoascorbate. We found that a single supplement of ascorbate and 2-phosphoascorbate adequately maintains intracellular vitamin C at physiological levels for up to 72 h.
Section of Biomedicine, Department of Disease Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg CDK-1870, Denmark.
Henriette
H
Frikke-Schmidt
Jens
J
Lykkesfeldt
D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D006801:Humans; D013997:Time Factors
C011669:ascorbate-2-phosphate; D001205:Ascorbic Acid