Ooops! I messed up! Help! - freezing cells (Dec/06/2007 )
I just realized that I really screwed up. I pelleted some human WBCs for freezing at -80. However, I forgot to discard the supernatant before freezing (supernatant consists of a PBS running buffer)!
What effect will this have on the cells? Will the viability be greatly effected when I'm ready to use them?
I'm confused! Do you usually freeze your cells as a pellet (ie not in any freezing medium)??
Yes, these particular cells are stored with no freezing medium. I will be using these cells fairly soon, so that's why no medium is used. For cells that will be stored longer, I use glycerol, or DMSO/FBS for liq. nit. storage.
The cells are gone as soon as you thaw them. Sorry to hear that. DMSO needs to be in there.
have you ever used this method of freezing cells before?
i have never heard of it. what were your cells, put through this method, like?
i always use DMSO freezing media. ice crystals are really sharp, and destroy in cells. DMSO prevents this.
Sorry, but your cells are gone. You definitely need DMSO as others also suggested.