Freezing PBMC for later analysis

From: Manetz, Scott (SManetz@Therimmune.com)
Date: Fri Dec 08 2000 - 09:35:41 EST


The lab I do work for has studies about to get underway in rats and
monkeys that have blood samples (into EDTA coated tubes) being taken at
multiple time points. We have always analyzed fresh blood, but these
studies are smaller and it would be better if we could wait until the
end to analyze everything instead of wasting time. I am interested in
spinning down the blood, removing the plasma, and then freezing the
remaining cells for analysis of PBMC subsets. My questions are:

1. We always used whole blood and lysed rbcs after staining, but can you
take a blood sample, spin it, take off the plasma, resuspend the pellet,
and still get the same values as if you had stained whole blood?
2. Should I just freeze the intact cell pellet after removing the plasma
or resuspend the pellet in media and then freeze?
3. If I need to resuspend in media, does it need to contain DMSO or be a
freezing media?
4. How long are the cells good for once frozen?

Any help would be greatly appreciated.

Scott Manetz



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