Comparison of the DNA methylation profiles of CD14+ monocytes from human peripheral blood with derived macrophages (MACs) and osteoclasts (OCs) obtained by exposure with M-CSF and M-CSF/RANKL, respectively, for 5 and 20 days

The methylation profiles of bisulfite(BS)- and oxidative-BS-modified DNA of human CD14+ monocytes were compared with derived macrophages (MACs) and osteoclasts (OCs) following M-CSF and M-CSF/RANKL incubation for 5 and 20 days, using the Infinium HumanMethylation450 BeadChips (Illumina, Inc., San Diego, CA,). This platform allows the interrogation of >485,000 methylation sites per sample at single-nucleotide resolution, and comprises an average of 17 CpG sites per gene in the 99% of RefSeq genes. 96% of CpG islands are covered, with additional coverage in CpG island shores and the regions flanking them. The samples were hybridized in the array following the manufacturer’s instructions. Total DNA isolated by standard procedures from CD14+ cells (total monocytes, MOs) corresponding to three sets of samples of monocytes (MOs) and derived MACs and OCs following incubation with M-CSF or M-CSF/RANKL for 5 and 20 days.