Hi all,
I am looking for a protocol detailing the optimal freezing and thawing conditions of PBMCs to ensure maximum viability and functionality of monocytes. I will be freezing down patient PBMCs and later thawing and analysing in batches, with the hope to examine T cells and monocytes. From what I can tell (and past experience) T cells are fairly robust and can survive this ordeal but monocytes have notoriously low survival and activity rates post-thaw. With no experience in working with monocytes, I wonder if anyone out there knows a certain temperature, media constituent or protocol that promotes monocyte survival and function.
Thank you in advance.
Christof
Hi Christof, I have only anecdotal reports of freezing back buffy coats of Ficoll separations.
B-cells and T-cells do OK, when you slowly freeze them in the Ficoll like you do for DMSO treated cells for LN2 storage.
Fast thawing is recommended, but platelets and neutrophils pose a problem. They don't like freezing at all.
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