freezing of hemopoietic stem cells
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
Where am I going wrong ?
Many thanks for any helpful tip.
Ralph M.Bohmer, New England Medical Center, Boston
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