I tried PMA + Ionomycin (conc 1ug/mL) and PHA (4 ug/mL) stimulation for 4Hrs. It didn't work. Any suggestions and ideas are welcomed.
U937 cells have a maturation block, they can be differentiated to macrophages or dendritic cells. Culturing the cells with 100ng/mL PMA for 48 hours significantly increase their macrophage like capabilities. Then they can be stimulated with LPS.
Right now, a member in my Lab, he's trying to use PMA to activate this cell line. with 100ng/ml . The best conditions are 0.5x10^6 cell/ml for 24 hours and 0.125x10^6 cells/ml for 48 hours in 24-well plate
In my previous lab which U937 comes from (Uppsala= U) and the patient was born in 1937. there are many publications about your question and I think this paper will giv you what you need.
http://www.ncbi.nlm.nih.gov/pubmed/11236937
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