We have successfully differentiated human-derived monocytes into M0 macrohages followed by cytokine polarization to achieve different macrophage phenotypes using traditional protocols with media supplemented with 10% FBS. We are interested in conducting these experiments using autologous human serum. I have seen protocols recommending purchased Human AB serum but that is not our desire. Does anyone have recommendations? Our primary concern is the presence of cytokines and growth factors already present which could interfere with downstream polarization experiments. Thanks for any input!
Hi Catherine,
In my opinion, the answer is no. I agree with your concern, serum proteins or cyokines modify diferentation process. In my experience, I used ascitic fluid to differentiate macrophages and M0 macrophages (only with FBS) were used as a control of changes in differentiation. I used the same proportion of ascitic fluid 10% and FBS 10% in RPMI.
I guess it depends why you want to use autologous serum- to go animal-free, or to understand how the patient serum modulates the macrophages? You could try adding HSA- human serum albumin- to your media, or use an optimised serum-free media such as X-VIVO 10 (which contains some HSA). I am using X-VIVO, but I am only just starting out so I can't yet tell you how it works for M0. For my M1/M2 macrophages, it works well. Otherwise, you could heat-treat your autologous serum to denature the cytokines, though this would obviously affect other proteins.
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