Purification of Normal Human Bone-Marrow-Derived Basophils

Abstract

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.

Histamine-releasing factor/translationally controlled tumor protein (HRF/TCTP)-induced histamine release is enhanced with SHIP-1 knockdown in cultured human mast cell and basophil models

“…However, transfection of human basophils and mast cells has proved extremely difficult, likely as a result of the fact that these cells are at an end-stage in their development. To circumvent this technical difficulty, we used culture-derived cells, and although many groups have described this for mast cells [20 -24], only a few investigators report the successful culture of basophils from precursor cells [25][26][27][28][29]. In this study, we show that sufficient numbers of basophils can be expanded from CD34ϩ cells that are suitable for transfection and signal transduction studies.…”

The identification and characterization of umbilical cord blood-derived human basophils

“…In most of these studies, progenitors in bone marrow or cord blood exposed to interleukin-3 (IL-3), interleukin-5, or a combination of both, give rise to a majority of eosinophils with basophils and basophil/eosinophil hybrids making up the next most abundant cell population. Although much has been learned from these cells, including evidence for a common basophil/eosinophil progenitor cell [15,[18][19][20][21][22], the conditions leading to the differentiation of a cell population containing predominately basophils have not been investigated thoroughly.…”

Establishment and characterization of continuous hematopoietic progenitors-derived pig normal mast cell lines

“…Indeed, several attempts have been made to induce the maturation of hematopoietic progenitors of different origin (cord blood, bone marrow or peripheral blood) in the presence of recombinant human IL-3. In all the reports published, including ours, such cultures have given rise to the differentiation of mixed basophil-and eosinophil-like cell populations (Arock et al, 1993;Valent et al, 1989). No MC development, even as a minor cell population, has been observed in this condition of culture.…”