Expired media - How long after the expiration data is cell culture media good? (Jun/15/2011 )

I have been trying to grow some cell lines with IMDM (Iscove's Modified Dulbecco's medium). The media is 6 months after the expiration date. Could it be the reason why the cells are growing very slowly? How long is media good after the expiration data? Thanks.

It depends on the medium formulation. Try adding some l-glutamine and see if that improves things. L-glut is one of the more labile of the compounds in most media.

distalless on Wed Jun 15 18:28:35 2011 said:

As Bob1 correctly states "it depends upon the formulation of the media"

It also depends upon what you want to do with the cells. Many years ago we had Gibco produce a media "special" for us. This was DMEM (1000mg/L Glucose) regular formulation BUT without L-Arginine. We wanted to grow our cells in L-Arginine free conditions and stimulate our cells to produce an inducible enzyme. As the "special" was expensive to purchase and we had to have a large minimum amount, this meant that we had plenty of media left over after our initial experiments........10 years later .......8.5 years after it's expiry date......the cells still grow, respire oxygen at a similar rate....and produce the inducible enzyme.

L-Glutamine should always be added to growth media...we purchase media always without Glutamine.....and then buy 100 X concentrated L-Glutamine. It is one of the most labile compounds added to media.


Some media needs to be kept in dark conditions in a cold room and can go of due to the light.


There are many other reasons why cells may grow slowly:

CO2 levels are either high/low.
Cellular Senescence due to high passage number
Contamination by mycoplasma's (as mycoplasma's cannot be seen under the phase contrast microscope)
Change of Serum batch.
Over Trypsinisation.

Hope this is useful.

Kindest regards.

Uncle Rhombus.