Hi everyone,
does anyone know the best condition to isolate and culture monocytes and monocyte-derived macrophages? Many papers suggest DMEM, others RPMI + 10% FBS. I am culturing these cells in RMPI+10% FBS for 2 weeks already. I started by observing WBCs (presumably monocytes), which eventually differentiated in macrophages (spindle-like and/or "fried egg" phenotype). Over the second week I noticed a decrease mostly in macrophages number and cell growth seems to have slowed down. I've searched for these specificities in the literature, but couldn't find anything specific to the duration of these cells in culture.
Any help would be much appreciated,
Best regards,
Luisa
One your cells are macrophages, you should work with them as soon as possible.
We grow human monocyte-derived macrophages using RPMI supplemented 10% human serum and use the cells usually between days 6-8. Hope this helps!
Hi Dearbhla,
it is usefuk. Thank you very much. Do you happen to know how long do monocytes/macrophages last in culture?
Macrophages are derived quickly from monocytes by using Phorbol Myristate Acetate (PMA), this happens with a period of overnight, thereafter you can store and revive your macrophages though it mustn't be for more than three months. With media RPMI supplemented 10% FBS will do.
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