I recently purchase some A431 cells from ATCC and the recommended media is DMEM + 10% FBS (my media is also supplemented with GlutaMAX). After thawing and passaging some cells attach, however the longer I leave them in culture more cells become detached. Anyone know why this could be? I have read that some people add more glutamine and amino acids.
Thanks!
with ATCC cells I had two times the experience that the cells were contaminated (it was a Sertoli-cell line). So this possibility is there, too.
Once I had problems with the incubator. Although the incubator CO2 display was okay, if measuring with an independent device the CO2, it came out that it was too low.
So after that the first thing I did if encountering strange things like this is to check CO2.
Many times the cells undergo stress whilst shipping and simply die dispite addition of extra nutrients...however i strongly advise you to check that ur incubator is correctly set and ur media is not outdated as cells get sensitive through passages. Also i would advise you to keep a little of the old media in the flask when subculturing as it contains growth hormones and other essential components that help it to grow
Thanks both for the suggestions. The addition of 2 mM L-Glutamine seems to have dramatically improved viability, which is surprising considering the media already contains GlutaMAX.
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