In Vitro Culture and Differentiation of Primary Monocytes into Macrophages

After counting, re-suspend the isolated monocytes at a 1 times 10 to the sixth cells per milliliter of 37 degrees Celsius serum-free RPMI 1640 medium supplemented with antibiotics, and plate 1 milliliter of re-suspend cells into 35-millimeter plates in a biosafety cabinet. Then, place the plates in the cell culture incubator for 30 to 60 minutes to allow the cells to adhere to the well bottoms.

To prime for macrophages with an M1-like phenotype, add 100 microliters of fetal bovine serum containing 25 nanograms per milliliter of GM-CSF to each plate. To prime for macrophages with an M2-like phenotype, add 100 microliters of fetal bovine containing 50 nanograms per milliliter of M-CSF to each plate.