Hi,
For example, the NK92-MI cell line, according to the manufacturer's instruction, 2-mercaptoethanol should be supplied as additional reagents. So my question is, what is the function of 2-mercaptoethanol in cell culture? Can it be replaced by glutathione?
Thanks!
beta- mercaptoethanol is a reducing agent used to stabilize certain culture media by avoiding the oxidation of certain unstable compounds? especially for very rich culture media. An example of stabilization is glutamine which is particularly unstable at 37 ° C. Beta-mercaptoethanol partially stabilizes glutamine, which is essential for the expansion of B cells. The same is true for certain vitamins, other amino acids and essential fatty acids.
I am interested as well to know the answer. I add 2-mercaptoethanol to bone marrow derived BMDC cultures and in vitro studies involve DCs and T helper cells co cultures.
2-Mercaptoethanol also known as beta-mercaptoethanol is a potent reducing agent used in cell culture media to prevent toxic levels of oxygen radicals. 2-Mercaptoethanol is not stable in solution, so most protocols require daily supplementation. Gibco 2-Mercaptoethanol contains 2-mercaptoethanol at a concentration of 55 mM in Dulbecco's phosphate buffered saline (DPBS).
As far as I understood, this is highly empiric.
Historically, β-mercaptoethanol (BME) has been shown to work well to culture lymphocytes (notably by a swiss immunologist called Theodor Brunner). More specifically, addition of BME to the cell culture medium resulted in about 30 fold increase of lymphocyte numbers in mixed leukocyte cultures, so the effect was considerable.
Whether the effect of BME is related to its reducing activity is highly unclear since BME is rather unstable and loses its reducing activity quite quickly upon incubation. Furthermore, it has been proven that reducing activity of BME was simply not necessary to mediate its effect (again, in relatively old studies in cell coculture systems that are not so much used nowadays). In some other cell cultures including non immune cells, it is seemingly used to avoid/remove toxicity, but I do not know whether or not it's related to BME's reducing activity.
Regarding your question about gluthatione, bare in mind that if you choose an approach that differs from the recommended one, you may expect reviewers (or colleagues) to ask you to show that your results are not influenced by this choice.
beta- mercaptoethanol is a reducing agent used to stabilize certain culture media by avoiding the oxidation of certain unstable compounds? especially for very rich culture media. An example of stabilization is glutamine which is particularly unstable at 37 ° C. Beta-mercaptoethanol partially stabilizes glutamine, which is essential for the expansion of B cells. The same is true for certain vitamins, other amino acids and essential fatty acids.
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