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Freezing and thawing primary lymphocytes - Any protocols? (Sep/26/2005 )

I need to freeze and thaw primary lymphocytes isolated from mouse blood and bone marrow. I would love some suggestions to increase my viable yield!!

THANKS!!! smile.gif



I have a little experience, here is what I have found...

According to the core lab where I get my cells, it is very important how the cells are frozen, they add the freezing medium drop by drop to cells already aliquotted to cryovials, they say drop by drop is essential!

Dilute each vial (for me 0.5-1*10^6 cells) into at least 10 ml of PRE-WARMED culture media do not put more than one thawed vial of cells per 10ml of media (again per the core facility)--the media must be pre-warmed to 37C so that the pH is just right for the cells....

Pellet gently, remove "wash" media and resuspend in culture media

Also check out some websites where they sell frozen primary cells like poetics the protocol for them is attached...