I immersed theTHP-1 cells with PMA at a concentration of 50 ng / ml for 72 hours and then rested them for 48 hours in a PMA free medium. But only a small percentage of the cells had become macrophages. And this number returns to the original state after a period of time. what should I do?
Dear Sara Molaghaee
Please check the attached step by step protocol from the Bowdish lab at McMaster University that I have used and adapted with great success.
http://www.bowdish.ca/lab/wp-content/uploads/2011/07/THP-1-propagation-culture.pdf
If you still have problems check the purity/identity of your THP-1 cell line through simple microsatellite profiling that ATCC or any good genomics facility will offer.
https://www.atcc.org/en/Services/Testing_Services/Cell_Authentication_Testing_Service.aspx#service
All the best & good luck with your experiments,
Michael
Dear @Michael Forster
Thank you so much, I really appreciate your help.
Base on my experience, you can't maintain differentiated THP-1 macrophage cells in media without the presence of PMA. They tend to revert back to monocyte cells slowly. And I use 200nM of PMA to differentiate THP-1 monocyte cells to macrophage-like cells for 72 hours. Sara Molaghaee
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