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293F cells revival - (Nov/24/2006 )

Has anyone faced problems reviving 293F cells. Recieved a new batch of cells but cells are unable to survive after thawing. Also, we do not spin down cells. Just transfer cells in a T25 flask with warm freestyle media (SFM). Does anyone has any insights of the possible problem with trying to grow the cells and any possible suggestions.

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-aqua26-

QUOTE (aqua26 @ Nov 24 2006, 05:57 AM)
Has anyone faced problems reviving 293F cells. Recieved a new batch of cells but cells are unable to survive after thawing. Also, we do not spin down cells. Just transfer cells in a T25 flask with warm freestyle media (SFM). Does anyone has any insights of the possible problem with trying to grow the cells and any possible suggestions.

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Probably these cells did not freeze right or have already went through a freez-thaw cycle prior to yours. Not much you can do at this point. Get another one from a diifefrent batch or from commercial source.

-genehunter-1-

Hi aqua26,
1) was the T-25 flask actually large enough? Sometimes I thaw cells in a T-75, because if they are too numerous for a small flask then they don't survive (personal experience).
2) try not only warming, but conditioning the media with CO2: I like to put my receiving flask of media in the incubator to get the CO2 levels up to snuff, then thawing my cells and adding them.
I also fully agree with genehunter-1: the cells may have been duff to begin with dry.gif
Good luck! biggrin.gif
mito1

-mito1-