Liquid nitrogen is the preffered way of long term storage of cells.
The - 80 C should really only be used for short term storage, so your cells should be perfectly fine when taken from liquid nitrogen
Thaw the cells from the liquid niotrogen very rapidly (in palm of your hand is best) as the DMSO is very toxic to the cells, so as soon as the last crystal of ice is gone place the cells in 1 ml of pre-warmed media and spin them down and remove the supernatant and resuspend the cells in fresh pre warmed media, this will remove all DMSO.
However,
You can also place them straight into the pre-warmed media (in a T25 or T75 for example) without centrifuging them and just place in the incubator to culture them as the DMSO will be diluted out, we work with HepG2 and never have any problems doing this.
Another tip is if you are losing many cells when you thaw them out (many float and do not survive you can increase the FBS to 10 % or even up to 20% to help them resucitate, just remeber to reduce this back to normal levels when the cells start growing well
After culturing for a few days you can then seed them for analysis
Cotchy.
Edited by cotchy, 19 March 2009 - 06:13 AM.
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