Hello,
Many protocols add b-mercaptoethanol to cell media (concentrations around 0.05 mM) in order to keep T cells, macrophages, and DC cultures viable; however, some protocols do not.
BME is a reducing agent and will completely change the redox environment of the cell culture if added at these concentrations and will also perturb drug interactions.
Since there are mixed protocols with some groups using it and some not I'm wondering if it is absolutely necessary? I believe my experiments will be more realistic, reliable, and informative in the absence of BME.
Best,
Hi Gregory,
Here's just some quick considerations. As you have pointed out people use 2-mercaptoethanol to increase viability in their cultures. In fact, it makes a huge difference especially when culturing mouse T cells with mitogenic stimuli like agonistic antibodies, PHA or PMA & ionomycin. It also influences T cell activation and proliferation, probably through altering the redox state of the cell and/ or the preferential use of glutamine over glucose as energy source. Now it totally depends on the type of experiments you want to do and the readout you plan to use. For a rather comprehensive overview, please see the attached review on effects of thiols on immune cells (Don't let the title fool you, they also provide a rather comprehensive reference list for murine cells).
Article Review: 2-Mercaptoethanol alteration of in vitro immune func...
All the best & good luck with your experiments,
Michael
Thank you Michael, I didn't know there was a review for this topic - very helpful!
Not necessary .. try to use RPMI media for immune cell harvisting
When I was first learning tissue culture I was told that 2-ME is very important for mouse cells and less so for human; something about mouse cells producing more free radicals than human cells in culture. I've always just gone with that, so I don't have a good literature source to point you to, but I just checked and this Purdue cytometry group post seems to mostly agree:
https://lists.purdue.edu/pipermail/cytometry/2002-April/022078.html
Hello
I agree with Michael Förster totally.
Article Use of 2-mercaptoethanol in cell culture
Good Luck
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line