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.
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.
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