L-glut in media? - (Oct/20/2008 )
I have another really stupid question. Since I'm using the Gibco DMEM which already has L-glut in it, do i have to add more L-glut into the media, or is 10% FCS alone will be enough? I'm worrying if im poisoning the cells with L-glut...
if your media already contained L-glutamine, I don't think you'll need to add in again.
i prefer to buy media without L-glutamine and add in myself. coz L-glutamine in media already added with it might become unstable if you store it after some times.
As far as I know when we bought medium with L-glutamine after 2 months we added a glutamine from stock to be sure that it was still present (because of its unstability). Now, we buy media without glutamine and we add it from stock.
Whether or not the glutamine is stable in storage and culturing depends on whether it is L-glutamine (degrades quickly into ammonia) or L-analyl-L-glutamine (stable in solution).
In our lab we use GlutaMAX DMEM (comes with L-analyl-L-glutamine), and have never added extra L-glut. Our cells grow big fat and healthy, and we've even used some bottles that were a few months past the best-before date with no noticeable change in cell growth.
See Invitrogen for more details.
Thanks for the input!
Will to much L-glut affect cell growth/vialibily?
I know L-glut will degrade to ammonia, so it will probably affect the pH of the media. I realize that my media in the dish was quite pink.
My cells are growing really slowly now, with lots of giant cells. I'm trying to find out what is the problem.
I did the same as you. I prepare the L-glutamine stock solution and stored at -20. I thaw it and use when I want to prepare DMEM complete media.
It is true that we have to add L-gln whether it is already there in the medium or the medium does not contain any L-gln b/c of its unstable character. My question is whether it is appropriate to discard the medium after 1 month if there is L-gln in it as I think it also cause toxicity to the cells because of the release of ammonia ? Is the bright pink color of the medium is because of this chemical change when the reconstituted complete medium is little old ?