I have it frozen already. So i will thaw them and resuspend in normal media 10 ml. Then I will add PMA in a conc of 1µl PMA to each 1ml medium. means for the 10 ml medium, I may add 10 µl PMA. then keep it overnight. then the THP-i now is macrophage and addhere to the surface. then I will trypsinize it, centrifuge. add new medium and then go ahead with culture subculture cycles
Sigma suggests a 20nM solution of DMSO to dissolve the PMA into, then once the PMA is dissolved dilute the mixture with water. DMSO is very cytotoxic to cells, so I would be careful adding DMSO at high concentrations.