Classical and non-classical monocytes are two well-defined subsets of monocytes displaying distinct roles. They differentially express numerous genes relevant to their primary role. Using five independent transcriptomic microarray datasets, we ruled out several inconsistent genes and identified common genes consistently overexpressed either in classical or non-classical monocytes. One hundred and eight genes were significantly increased in classical monocytes and are involved in bacterial defense, inflammation and atherosclerosis. Whereas the 74 genes overexpressed in non-classical monocytes are involved in cytoskeletal dynamics and invasive properties for enhanced motility and infiltration. These signatures unravel the biological functions of monocyte subsets. HIGHLIGHTS We compared five transcriptomic GEO datasets of human monocyte subsets. 108 genes in classical and 74 genes in non-classical monocytes are upregulated. Upregulated genes in classical monocytes support anti-bacterial and inflammatory responses. Upregulated genes in non-classical monocytes support patrolling and infiltration functions.