freezing PBMC - (Nov/03/2006 )
do anybody have any experience with freezing of PBMCs?
yes, after Ficoll freeze pellet in FCS/DMSO 9:1
no problem after thawing for dendritic cell production
For lymphocytes I do medium with 50% FCS and 10 % DMSO
It is very important that the DMSO is introduced slowly. I add medium with 20% DMSO to an equal volume of medium & cells to obtain the 9:1 ratio. You also want to do this on ice; as, the DMSO tends to heat things up a bit. Put cells in a controlled-rate freezing environment immediately!
I use 70% FCS, 20% RPMI and 10% DMSO. Important to freeze in a temperature controlled environment (i.e. one of those boxes with isopropanol underneath)