Column separation of monocytes by adherence to gelatin beads

This investigation was performed to study whether the efficient binding of collagen to monocytes in the presence of fibronectin and heparin may be used for separation of monocytes from human peripheral blood. It was shown that monocytes adhere selectively to gelatin bead columns in the presence of fresh plasma and heparin. Mononuclear blood cells are rapidly depleted of monocytes by passage through a 5-10 ml column at a flow rate of 1.5-2.0 ml per min. Adhering lymphocytes are more loosely attached and may be detached by stirring and washing, while the monocytes can be eluted by 50 mM EDTA. This separation technique is suitable for combination with various other methods since it is rapid, allows convenient handling of large numbers and yields cells with very high viability. Although most B lymphocytes pass through the column without attaching, there is some enrichment of B cells and non-T, non-B cells among the adherent lymphocytes.