Macrophage Differentiation Using M-CSF

Results Summary

Here, we describe the differentiation of macrophages from monocytes isolated from PBMCs. After PBMCs, extraction, monocytes were isolated using the Pan Monocyte Isolation Kit II (see different review). Then, monocytes were cultivated in RPMI supplemented with 20% FCS, HEPES and 1%Glutamin. Then, M-CSF was added at a final concentration of 50ng/ml. After differentation for 3 d, the supernatant was removed carefully, cells were washed carefully with media once and media supplemented with M-CSF at a final concentration of 50ng/ml was added again. After 3d, (6d overall), the cells should have phenotypically changed as observed by microscopy and should be differentiated into M2 macrophages.