hi everyone,

im trying to study macrophages polarization in critically ill patients. Unfortunately, the only samples I have are biobanked plasma samples. I am trying to treat commercially available naive monocytes with the patient plasma sample to induce macrophages polarization (since theoretically the cytokines concentration in the plasma sample should drive M1/M2 polarization ratio reflective of what was in the original blood sample). Can anyone suggest a protocol for the mentioned experiment? What is the naive monocytes count I should start with, media to grow the cells and volume of patient plasma that I should treat the monocytes with to induce polarization, and duration of the culture. I am planning on quantifying M1 and M2 macrophages in the resulting culture using flow cytometry. Looking forward to seeing your feedback. Thanks.