Cryopreserved Dendritic Cells?

Wijewardana, Viskam viskam at pitt.edu
Thu Aug 7 09:02:44 EDT 2008


Hi Mario,
In our laboratory we use cryopreserved Rhesus PBMC, lymph node and spleen suspensions to
enumerate DCs for studies. Once thawed they have more than 90% viability. However during
culture they undergo increased death. This could be rescued by adding GMCSF, IL-4 and
IL-3 to the cultures. Again this survival is more pronounced in myeloid DCs compared to
Plasmacytoid DCs.
So, yes, we do agree with what you have observed not otherwise. I guess the thawing
conditions are critical in getting DCs back from frozen samples.
Any comments?
-Viskam

Viskam Wijewardana
Barrat Boyes Lab
Center for Vaccine Research
9051 Biomedical Sciences Tower 3
University of Pittsburgh
3501 Fifth Avenue
Pittsburgh, Pennsylvania 15261
Ph 412 383 9057
Fax 412 624 4577
________________________________________
From: Mario Roederer [roederer at nih.gov]
Sent: Tuesday, August 05, 2008 8:17 PM
To: cyto-inbox
Subject: Cryopreserved Dendritic Cells?

There is a persistent lore that you cannot cryopreserve dendritic
cells in PBMC -- that if you freeze PBMC and thaw them, you don't get
the DC's back out.  Recently, we were comparing fresh vs. frozen PBMC,
and did not observe this (if anything, the frozen PBMC had more
DC's).	However -- this was done only phenotypically and not
functionally.

Can anyone point me to data that shows that DC's do not survive
cryopreservation?  Are there references to this?

mr

_______________________________________________
Mario Roederer, Ph.D.
Senior Investigator, ImmunoTechnology Section
Vaccine Research Center, NIAID, NIH
40 Convent Dr., Room 5509
Bethesda, MD 20892-3015
Phone: 301 594-8491
FAX: 301 480-2788




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