BACKGROUND: The immunomagnetic separation technique is the basis of monocyte isolation and further generation of monocyte-derived dendritic cells. OBJECTIVE: To compare the efficiency of monocyte positive and negative separation, concentration of beads, and their impact on generated dendritic cells. METHODS: Monocytes were obtained using monoclonal antibody-coated magnetic beads followed the Ficoll-Paque gradient separation of mononuclear cell fraction from the peripheral blood of 6 healthy volunteers. CD14 expression was analyzed by flow cytometry. CONCLUSIONS: Both types of magnetic separation including recommended and reduced concentrations of beads did not affect the yield and the purity of monocytes and their surface CD14 expression. However, DCs originated from the "positively" separated monocytes had noticeable higher expression of CD80.
BACKGROUND: The immunomagnetic separation technique is the basis of monocyte isolation and further generation of monocyte-derived dendritic cells. OBJECTIVE: To compare the efficiency of monocyte positive and negative separation, concentration of beads, and their impact on generated dendritic cells. METHODS: Monocytes were obtained using monoclonal antibody-coated magnetic beads followed the Ficoll-Paque gradient separation of mononuclear cell fraction from the peripheral blood of 6 healthy volunteers. CD14 expression was analyzed by flow cytometry. CONCLUSIONS: Both types of magnetic separation including recommended and reduced concentrations of beads did not affect the yield and the purity of monocytes and their surface CD14 expression. However, DCs originated from the "positively" separated monocytes had noticeable higher expression of CD80.