Primary Macrophage polarization
Description
Primary macrophage polarization on 3D printed PCL scaffolds normalized to M0 on TCP
Files
Steps to reproduce
To generate macrophages, culture primary CD14+ monocytes for 5 days with RPMI 1640 supplemented with 10 % heat-inactivated human serum in the presence of 20 ng/mL of the maturation factor, hM-CSF in a 24 well plate. For polarization, macrophages were incubated in RPMI medium supplemented with lipopolysaccharide (PLS) and IFN-γ at two concentrations (100 ng/ml LPS and 10 ng/ml IFN-γor 250 ng/ml LPS[8] and 20 ng/ml IFN-γ. Potential M2 macrophages were incubated in RPMI medium containing 20 ng/ml IL-4. Positive control samples (M0-like macrophages) were cultured in medium without any cytokines. After 48 hours of stimulation, cells were harvested for gene expression or fluorescent analysis.