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cell culture - (Nov/30/2011 )

I am using PBMC for my experiment with monocytes which has to be converted into dendritic cell. I tried to preserve the PBMC at -80C with 10% DMSO and 90% Heat inactivated serum. After one month i checked the viability of monocytes on flow cytometry with Antibody CD14 and 7AAD which has come closer to 60-65%.
When i plated this on my nunc surface treated plates for 2-3 hours , then monocytes are not attaching to the surface,almost 95% of the cells are non adherent. Could anybody help me with this? though i am going to try one monocyte attachment medium just launched by Promocell. I am not sure whether it will help to solve my problem or not.
Please help.


hi, First of all i would like to ask you a few questions:

(1) what is PB?? (PBMC)
(2)why should monocyte be converted to dendritic cell

Yes, i have used one such specific media from Promocell for myocytes and they were good - in fact they also have specific trypizining media and freezing media, which are worth trying to save the cells.

-materials researcher-