Purification of morphologically and functionally intact human basophils to near homogeneity.
Certain studies on basophils require highly purified functionally intact cell preparations. Here, a three-step procedure is described that meets these requirements. The procedure consists of a Ficoll density gradient step, counterflow elutriation and negative selection by magnetic cell sorting (MACS). The mean purity of basophils obtained from 30 donors was 97.6+/-3.96% with a viability of 99.6+/-0.83%. The recovery rate was 49.7+/-15.6%. The cells had a normal morphological appearance as assessed by May-Grünwald-stained cytospins and were functionally intact as shown by their unaltered capacity to release histamine and interleukin 4 (IL-4) following immunological activation. This procedure is a clear improvement over currently available techniques and should facilitate future investigations on basophils.
Haisch
K
K
Division of Cellular Allergology, Research Center Borstel, Germany.
Gibbs
B F
BF
Körber
H
H
Ernst
M
M
Grage-Griebenow
E
E
Schlaak
M
M
Haas
H
H
D001491:Basophils / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D015847:Interleukin-4 / Q000096:biosynthesis
D015847:Interleukin-4; D007073:Immunoglobulin E