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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?

Thanks!

-Yasy-

I'm confused! Do you usually freeze your cells as a pellet (ie not in any freezing medium)??

-lauralee-

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.

-Yasy-

The cells are gone as soon as you thaw them. Sorry to hear that. DMSO needs to be in there.

-genehunter-1-

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.

V

-vetticus3-

Sorry, but your cells are gone. You definitely need DMSO as others also suggested.

-scolix-