Hello all,
I am isolating fresh PBMCs from healthy donors to do some gene expression studies after infection with viruses or stimulation with TLRs agonist. My question is, can I keep the isolated PBMCs in culture medium for 24 hour then I can start the experiment? or should I work with it just direct after the isolation to avoid any influence might happen due to this 24 hour incubation?
Thanks in advance.
Yes start immediately after isolation because many cell types in PBMCs wont survive for longer times.
Thanks Goodwin for your answer. If I am interested only in monocytes, Will not this type of cell population survive for 48 hr?
It might. But since you want to study gene expressions, the earlier, the better..!!
hi mohamed, you need to start immediately. monocytes may attach, change their activation status and early stages of macrophage differentiation may occur. are you isolating your moncytes after the stimulation procedure?
Hi Pia,
thanks for your input, Actually I am interested to check the expression of one gene which is exclusively expressed in monocytes, so I got a recommendation to leave the PBMCs setting for 24 hr, then remove the unattached cells then you will get the monocytes adherent and ready for infection afterwards..
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