freezing of hemopoietic stem cells

Ralph Bohmer ralph.bohmer at es.nemc.org
Mon Oct 5 08:57:04 EST 1998


I am trying to freeze mononuclear cell preparations from peripheral blood, to 
use for colony cultures. After thawing, I get no colonies. Everything dead. 
(I get plenty of colonies before freezing). 
I freeze by suspending the cells in PBS or RPMI with 10-30% serum and 7-10% 
DMSO, then use controlled-rate freezing device (from Nalgene) to bring down 
to -80C, then nitrogen. Thawing by slowly diluting thawed suspension with 
room temperature PBS/BSA, then wash. Works wonderfully for cultured 
fibroblasts.

Where am I going wrong ?

Many thanks for any helpful tip.

Ralph M.Bohmer, New England Medical Center, Boston



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