Isolation and maintenance of nonadherent quiescent human monocytes for studies of adhesion and migration

A method is described for the isolation and culture of large numbers of nonadherent quiescent human peripheral blood monocytes for adhesion/migration studies. Approximately 1.0×10⁸ cells/white cell concentrate with > 95% viability are isolated by countercurrent centrifugal elutriation. These cells are initially activated by the isolation procedure, but after a minimum culture time post isolation of 48 hours and a rest period after radiola-belling of at least 3 hours quiescent cells are routinely obtained, with low basal levels of adhesion and a rapid, reproducible superoxide response to activation with phorbol myristate acetate.