I am using mammalian cell cultures to express a protein. Which concentration of Vit C should I use to add in the culture to "help" my mammalian cells?
Hello Tany Nino
Ascorbic acid when added to culture medium is not readily available to cells in culture because it is unstable in aqueous media. Ascorbate levels can be maintained in cell culture media by frequent addition of vitamin C, but the persistent oxidation will continuously generate dehydroascorbic acid. Extracellular concentrations of dehydroascorbic acid are considered non-physiological. Even the L-ascorbic acid in commercially available culture media, which contain the vitamin in their formulations, may be available for the cells in culture far lower than prescribed.
Culturing cells with vitamin C requires control over many aspects of the media and culture conditions. Monitoring ascorbate levels and limiting oxidation may not be sufficient to fully recapitulate the physiological roles of vitamin C.
The stability of ascorbate can be enhanced by low oxygen growth conditions, and the use of stabilized derivatives of ascorbate such as ascorbate-2-phosphate (AAP) that cannot participate in redox chemistry outside the cell, yet can maintain physiological intracellular ascorbate levels.
A mixture of 0.25 mmol/L ascorbate and 0.45 mmol/L ascorbate-phosphate would provide a constant concentration of ascorbate in the culture medium. This constant ascorbate concentration is nontoxic for cells and may stimulate cell growth in the short term and long term.
You may want to refer to the article attached below.
https://pubmed.ncbi.nlm.nih.gov/11249202/#:~:text=A%20mixture%20of%200.25%20mmol,short%20term%20and%20long%20term.
Best.
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