Flow cytometric determination of basophils in whole blood with n-propyl Astra Blue lodide

Malin, Michael J.; Hwang, Deng R.; Ben-David, Daniel

doi:10.1016/0003-2697(86)90145-4

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…Basophils exhibit a density lower than 1.077 g/ml and therefore usually remain in the mononucle ar cell fraction in contrast to neutrophils and eosinophils. Unfortunately, given the fact that the mean percentage of basophils in circulating white blood cells in healthy adults ranges from 0 to 2% (mean: 0.3-0.6%) or 10-80 cclls/mm' [21,22], the various techniques used to directly purify hu man basophils from peripheral blood cells allow one to ob tain no more than 5 • 106 nonactivated purified melachromatic cells, an insufficient yield for the biochemical and pharmacological analysis of these cells at the molecular level.…”

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Purification of Normal Human Bone-Marrow-Derived Basophils

Arock¹,

Mossalayi²,

Goff³

et al. 1993

Int Arch Allergy Immunol

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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-A 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.

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confidence: 99%