Hi, 

Your question was not clear. 

But however, you differentiate THP-1 into macrophages using PMA. It can be used at different concentrations titrating for the time. In my experience incubating THP-1 with 20 ng/mL of PMA for 24 hr followed by wash and further incubation for 48 hr worked fine. But you can titrate the amount and time as per your conditions and follow up experiment. If you want to use higher concentration of PMA then reduce the the time. you need not give rest of 48 hr. This all depends on what your experiment is? Again The cell number and media volume depends on what your experiment is and also the cell culture dish or plate you will be using.

For Mycobacterial infection, the MOI depends on cells and also the bacterial strain you are using. Typically for Mtb, the MOI generally used is in the range of 1 to 10 bacteria per cell. MOI has to be chosen such that it does not affect the cell death over infection time. There are some papers where they have higher MOIs. 

For example, in a 6 well dish, I seed around 1.2-1.5 million cells with PMA (20 ng/mL) and differentiate for 24 hr and rest for 48 hr. Then infect with Mtb (MOI is your choice).

You will find many papers on all these by simply putting keywords such as THP-1 and PMA or THP-1 and Mtb in either PubMed or Google. 

Hope this helps.

Hi Rakesh, thank you so much for your feedback. hopefully, it will help a lot.

Hi

Look at this link and PDF attached

hope help you

Hi Awatif,

Thank you so much for your nice cooperation.