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Mast cells cultured from the peripheral blood of normal donors and patients
with mastocytosis originate from a CD34+/Fc epsilon RI- cell population
M Rottem, T Okada, JP Goff and DD Metcalfe
Allergic Diseases Section, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, MD 20892.
Mast cells may be cultured from human peripheral blood in the presence of
recombinant human stem cell factor (rhSCF). The characteristics of the
cells in peripheral blood that give rise to mast cells are unknown. Because
mast cell precursors in human marrow are CD34+, human peripheral blood
mononuclear cells from patients with mastocytosis and normal controls were
sorted on the basis of CD34 expression and the positive and negative cell
populations were cultured in rhSCF, recombinant human interleukin-3
(rhIL-3), or both for 6 weeks. Cell cultures were examined every 2 weeks
for total and mast cell number and cell differential using Wright Giemsa
and acid toluidine blue stains and antibodies to mast cell tryptase and
chymase, cell-associated histamine, and expression of CD34, c-kit, Fc
epsilon RI, and Fc gamma RII using flow cytometric analysis. The
ultrastructural anatomy of mast cells was examined by electron microscopy.
Peripheral blood CD34+ cells cultured in rhSCF with or without rhIL-3 gave
rise to cell cultures consisting of greater than 80% mast cells by 6 weeks.
CD34+ cells cultured in rhIL-3 alone did not give rise to mast cells,
whereas rhIL- 3 plus rhSCF increased the final mast cell number eightfold
when compared with cells cultured in rhSCF alone. Mast cells increased
concomitantly with a decrease in large undifferentiated mononuclear cells.
CD34- cells did not give rise to mast cells. Histamine content per cell at
6 weeks was approximately 5 pg. Electron microscopy of 4- week cultures
showed immature mast cells containing predominantly tryptase-positive
granules that were either homogeneous or contained lattice structures,
partial scroll patterns, or central dense cores and mixtures of vesicles,
fine granular material, and particles. The CD34+ population at day 0
expressed Kit (65%) and Fc gamma RII (95%), but not Fc epsilon RI, by
fluorescence-activated cell sorter analysis. At 6 weeks, CD34+-derived mast
cells exhibited Fc epsilon RI in addition to Kit and Fc gamma RII, and were
negative for CD34 antigen. Patients with mastocytosis showed a higher
number of mast cells per CD34+ cell cultured compared with normal controls.
Thus, the mast cell precursor in human peripheral blood is CD34+/Fc epsilon
RI- and gives rise to mast cells in the presence of rhSCF with or without
rhIL-3, and the number of mast cells arising per CD34+ cell in culture is
greater when the CD34+ cells are obtained from patients with mastocytosis
compared with normal subjects.
Volume 84,
Issue 8,
pp. 2489-2496,
10/15/1994
Copyright © 1994 by The American Society of Hematology
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