Keeping the intracellular vitamin C at a physiologically relevant level in endothelial cell culture

TY - JOUR

T1 - Keeping the intracellular vitamin C at a physiologically relevant level in endothelial cell culture

AU - Frikke-Schmidt, Henriette Rønne

AU - Lykkesfeldt, Jens

PY - 2010/2/1

Y1 - 2010/2/1

N2 - 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 μM 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.

AB - 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 μM 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.

U2 - 10.1016/j.ab.2009.09.040

DO - 10.1016/j.ab.2009.09.040

M3 - Journal article

C2 - 19782654

SN - 0003-2697

VL - 397

SP - 135

EP - 137

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 1

ER -