title: Characterization of 'adult-type' mast cells derived from human bone marrow CD34(+) cells cultured in the presence of stem cell factor and interleukin-6. Interleukin-4 is not required for constitutive expression of CD54, Fc epsilon RI alpha and chymase, and CD13 expression is reduced during differentiation.
abstract: BACKGROUND
  In vitro-derived human mast cells exhibit different properties, depending in part on the source of progenitor cells. Most investigations have used fetal liver, cord blood or peripheral blood. Few have used adult bone marrow.
  
  
  OBJECTIVE
  Human mast cells derived in vitro from the CD34(+) progenitors in bone marrow and cord blood that had been cultured with recombinant human stem cell factor (rhSCF) and recombinant human interleukin-6 (rhIL-6) were compared.
  
  
  RESULTS
  After 12 weeks of culture, nearly all of the cells were mast cells, and nearly all of these had cytoplasmic granules containing both tryptase and chymase (MCTC type), stained metachromatically with acidic toluidine blue, and expressed CD117 on the cell surface. Both tryptase protein and mRNA were detected by two weeks of culture. Chymase mRNA and protein were detected at 4 weeks but not at 2 weeks of culture. By 12 weeks, chymase content per cell, measured by ELISA, was significantly higher (P < 0.05) in human bone marrow-derived mast cells (HBMMC) (5.6 +/- 0.9 pg) than in cord blood-derived mast cells (CBMC) (2.4 +/- 0.9 pg), whereas histamine and tryptase levels were not significantly different. Of the cluster designations tested, CD29, CD49d, CD51 and CD61 were strongly expressed on HBMMC. CD54 and Fc epsilon RI alpha also were expressed constitutively. Approximately half of CD34-sorted cells at day 0 were CD13(+) and this diminished as mast cell maturation occurred. Electron microscopy revealed that 12-week-old HBMMC had many secretory granules that contained spherical electron dense cores surrounded by electron lucent space, consistent with previous reports of immature MCTC cells developing in vivo.
  
  
  CONCLUSIONS
  CD34(+) progenitors of human bone marrow are a rich source of mast cell progenitors capable of expressing granule and surface markers of mature mast cells in the presence of rhSCF and rhIL-6.
authors.affiliation: Department of Internal Medicine, Gunma University School of Medicine, Maebashi, Japan. yshimizu@med.gunma-u.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Shimizu
authors.forename: K
authors.initials: K
authors.lastname: Sakai
authors.forename: T
authors.initials: T
authors.lastname: Miura
authors.forename: T
authors.initials: T
authors.lastname: Narita
authors.forename: H
authors.initials: H
authors.lastname: Tsukagoshi
authors.forename: Y
authors.initials: Y
authors.lastname: Satoh
authors.forename: S
authors.initials: S
authors.lastname: Ishikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Morishita
authors.forename: S
authors.initials: S
authors.lastname: Takai
authors.forename: M
authors.initials: M
authors.lastname: Miyazaki
authors.forename: M
authors.initials: M
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Saito
authors.forename: H
authors.initials: H
authors.lastname: Xia
authors.forename: L B
authors.initials: LB
authors.lastname: Schwartz
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis* / Q000187:drug effects*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D053818:Chymases; D006801:Humans; D007150:Immunohistochemistry; D015847:Interleukin-4 / Q000494:pharmacology*; D015850:Interleukin-6 / Q000494:pharmacology*; D008407:Mast Cells / Q000145:classification* / Q000166:cytology* / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000187:drug effects* / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology*; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism; D053802:Tryptases
chemical_list: D015703:Antigens, CD; D015850:Interleukin-6; D012333:RNA, Messenger; D019089:Stem Cell Factor; D015847:Interleukin-4; D012697:Serine Endopeptidases; D053818:Chymases; D053802:Tryptases