title: Phenotypic and functional characterization of in vitro cultured human mast cells.
abstract: Mast cell progenitor cells, derived from CD34+hematopoietic stem cells, enter the circulation and subsequently mucosal or connective tissues where they mature to mast cells. Upon activation, mast cells increase the expression of activation markers, e.g. CD63, and release histamine amongst other mediators. Traditionally, release of these mediators is quantified using assays measuring their extracellular concentration in the supernatant of stimulated cells.
  
  
  
  Human mast cells (HuMC) were cultured from peripheral blood, phenotypically characterized, passively sensitized with allogenic IgE antibodies and finally stimulated by anti-IgE that crosslinks IgE/FcεRI complexes. Alterations in the number of cells positive for CD63 and release of histamine were quantified simultaneously by flow cytometry.
  
  
  
  In culture, two distinct CD45+cell populations were identified: CD117+CD203c+hiand CD117-CD203c+lowcells. Both populations showed positivity for FcεRI, tryptase and chymase, and contained histamine. Activation resulted in a significant increase of cells positive for CD63+up to 21% (range: 11-39) for CD117+CD203c+hicells (P = 0.005), and 27% (18-55) CD63+for CD117-CD203c+lowcells (P = 0.02). Baseline histamine content was higher for CD117+CD203c+hicells than for CD117-CD203c+lowcells, respectively 994 (695-6815) Molecules of Equivalent Specific Fluorochrome V500 per cell (MESF-V500/cell) and 797 (629-4978) MESF-V500/cell (P = 0.02). After activation, CD117+CD203c+hicells showed significant histamine release of 578 (366-1521) MESF-V500/cell, whilst CD117-CD203c+lowcells resulted in 310 (217-366) MESF-V500/cell histamine release.
  
  
  
  This study discloses that culturing HuMC from CD34+progenitors yields 2 phenotypically distinct cell populations that display a greatly similar response upon cross-linking of IgE/FcεRI complexes. © 2016 International Clinical Cytometry Society.
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Cop
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: I I
authors.initials: II
authors.lastname: Decuyper
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: M A
authors.initials: MA
authors.lastname: Faber
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Sabato
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: C H
authors.initials: CH
authors.lastname: Bridts
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.|Department of Pediatrics, Laboratory of Experimental Medicine and Pediatrics, Faculty of Medicine and Health Science, University of Antwerp, Antwerp, 2610, Belgium.
authors.forename: M M
authors.initials: MM
authors.lastname: Hagendorens
authors.affiliation: Department of Pediatrics, Laboratory of Experimental Medicine and Pediatrics, Faculty of Medicine and Health Science, University of Antwerp, Antwerp, 2610, Belgium.
authors.forename: B Y
authors.initials: BY
authors.lastname: De Winter
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: L S
authors.initials: LS
authors.lastname: De Clerck
authors.affiliation: Department of Immunology, Allergology, Rheumatology, Faculty of Medicine and Health Science, University of Antwerp, Antwerp University Hospital, Antwerp, 2610, Belgium.
authors.forename: D G
authors.initials: DG
authors.lastname: Ebo
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006632:Histamine / Q000378:metabolism*; D006636:Histamine Release / Q000276:immunology; D006801:Humans; D008407:Mast Cells / Q000166:cytology*; D010641:Phenotype
chemical_list: D000888:Antibodies, Anti-Idiotypic; C026408:anti-IgE antibodies; D006632:Histamine
keywords: IgE; flow cytometry; histamine; mast cell activation