The Application of Dextran Sedimentation as an Initial Step in Neutrophil Purification Promotes Their Stimulation, due to the Presence of Monocytes

Figure 2

Dextran sedimentation prior to Ficoll-Hypaque density gradient centrifugation induces differences in neutrophil function compared to the 1-step method. Neutrophils were purified from blood by 1-step or 2-step with dextran sedimentation prior to low-density Ficoll-Hypaque gradient centrifugation. These cells were then applied to assays of (a) adherence (), (b) chemotaxis (), (c) Staphylococcus aureus killing (), and oxidase burst by (d) DHR-123 reduction () and (e) lucigenin-dependent chemiluminescence (). In (d), a representative overlay histogram of reduced DHR-123 fluorescence is shown, whilst in (e), chemiluminescence kinetics plots for three individuals, A, B, and C, are presented, with a graph of the comparison between the peak RLU.

(a)

(b)

(c)

(d)

(e)