Induction of differentiation of human mast cells from bone marrow and peripheral blood mononuclear cells by recombinant human stem cell factor/kit-ligand in long-term culture

Abstract

In the murine system, a number of cytokines (including interleukin-3 [IL-3], IL-4, and stem cell factor [SCF]) promote the growth of mast cells (MCs). However, so far little is known about factors controlling differentiation of human MCs. Recent data suggest that human MCs express receptors (R) for SCF. The aim of the present study was to investigate whether recombinant human (rh) SCF induces differentiation of human MCs from their precursor cells. For this purpose, bone marrow (BM; normal donors, n = 6) and peripheral blood (PB; normal donors, n = 11) mononuclear cells (MNC) were cultured in the presence of rhSCF, rhIL-3, rhIL-4, rhIL-9, recombinant human macrophage colony-stimulating factor (rhM-CSF), or control medium in long-term (8 weeks) suspension cultures. After 4 weeks, up to 5% of the MNC (BM and PB) cultured in the presence of rhSCF, but not in the presence of other cytokines, were found to exhibit the characteristics of MCs. These MCs expressed the YB5.B8-reactive domain of the SCF R as well as IgE R, as determined by combined toluidine blue/immunofluorescence staining. Myeloid antigens, likewise expressed on human basophils (ie, CD11b, CDw65, and Bsp-1), could not be detected on these cells. Furthermore, rhSCF, but not rhIL- 3, rhIL-4, rhIL-9, or rhM-CSF, induced dose- and time-dependent increases in the formation of cellular tryptase (an MC-specific enzyme) (rhSCF [100 ng/mL], 1,308 +/- 679 ng/mL v control medium, 18 +/- 6 ng/mL tryptase on day 35 of PB cell cultures), as well as an increase in cellular histamine. After 6 to 8 weeks, when other mature hematopoietic cells decreased, MCs still could be detected in culture, with up to 40% of all cells being MCs. To test whether rhSCF also activates tissue MCs, we performed histamine release experiments (dispersed tissue; lung, n = 3; uterus, n = 3). SCF was found to enhance (by up to 3.4-fold) the capacity of the MCs to release histamine upon cross-linkage of IgE R with anti-IgE. Together, these observations suggest that rhSCF induces in vitro differentiation of human MCs from their BM and PB precursor cells in long-term culture and upregulates MC releasability.

Nerve growth factor‐β induces mast‐cell marker expression during in vitro culture of human umbilical cord blood cells

“…HS instead of FCS, 61 may be a possible explanation for our current ®ndings. Since both murine and human NGF have been shown to be biologically active in basophils 62 and since interspecies DNA sequences are highly homologous, 1 the use of murine NGF in the studies of Valent et al 61 are on the other hand unlikely to account for the negative ®ndings of these authors.…”

Interferon‐γ promotes the survival and FcεRI‐mediated histamine release in cultured human mast cells

“…23,24 On the other hand, SCF enhances histamine release from human mast cells after a brief incubation. [28][29][30] Probably, the mechanisms of the IFN-g-and SCF-induced enhancement of histamine release differ.…”

Mast cell/fibroblast interactions

“…Nocka et al isolated a growth factor from conditioned medium of Balb/3T3 fibroblasts which had the biological properties expected of SCF [19]. It was shown that long-term incubation with SCF led to differentiation of human and murine MC from their precursor cells in the absence of fibroblasts [20,21]. In vivo, local administration of SCF promotes CTMC development in mouse skin, while its systemic administration in rats induces both CTMC and MMC development [22].…”