I have done THP-1 cell differentiation into macrophages. I used RPMI as a media. and PMA 30nM concentration. after 24 hours incubation there were many floating cells. what could be the reason? and what is the effect of it to differentiation process.
Hi
it depends on the passage number and conditions of THP-1 which determines how efficiently they can be differentiated.
how old is your PMA,was it freeze thawed many times..these factors also effect the differentiation process.
you also check the cell culture plate you are using..try to use cell-bind or something similar.
Thank you so much. PMA I used are freezes and thaw few times and THP-1 cell passage number was also high so I guess that was the reason. Thank you for your useful answer.
Stop using your THP-1 after about 2 months and thaw a new stock. Phenotypic changes in old THP-1 will prevent them from differentiating efficiently, as Atoshi mentioned. You should also aliquot your PMA to reduce freeze/thaw cycles, but I suspect this is less the issue.
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