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.
1 reply to this topic
#1
Posted 30 November 2011 - 09:23 PM
#2
Posted 01 December 2011 - 07:10 PM
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.
regards
(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.
regards
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