Purification of normal human bone-marrow-derived basophils.
It is well established that basophils and eosinophils share a common differentiation pathway, although the factors regulating their terminal commitment (towards one or other lineage) are not yet fully defined. Interleukin-3 (IL-3) is a major differentiation factor for both human eosinophils and basophils, yielding a mixed population composed predominantly of eosinophilic cells (65 +/- 9%; n = 4), basophils at different stages of maturity (29 +/- 6%; n = 4) and monocytes/macrophages (6 +/- 3%; n = 4), after 3-4 weeks in culture. Using a relatively rapid and simple method involving a first step of gradient density centrifugation over a Percoll gradient (d = 1.063 g/ml) and a subsequent step of adhesion on tissue-culture-treated plastic, a cell population composed of 94 +/- 5% normal basophils and their precursors, with no demonstrable mast cells, was reproducibly obtained from human hematopoietic cells cultured for 3-4 weeks in the presence of recombinant IL-3. These cells contained high levels of histamine (1.39 +/- 0.14 pg/cell) and released this mediator upon stimulation with calcium ionophore A23187 and in a dose dependent manner upon stimulation with IgE-anti IgE, demonstrating their functional capacity. This relatively simple method therefore permits the production of large quantities of pure populations of normal and functional human bone-marrow-derived-basophils.
Arock
M
M
Laboratoire d'Hématologie, Faculté de Pharmacie de Paris V, France.
Mossalayi
M D
MD
Le Goff
L
L
Dugas
B
B
Mencia-Huerta
J M
JM
Debré
P
P
Guillosson
J J
JJ
D001491:Basophils / Q000166:cytology* / Q000276:immunology; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007377:Interleukin-3 / Q000276:immunology; D011205:Povidone; D011994:Recombinant Proteins; D012822:Silicon Dioxide
D007377:Interleukin-3; D011994:Recombinant Proteins; D007073:Immunoglobulin E; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone