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Molecular and stimulus-response profiles illustrate heterogeneity between peripheral and cord blood-derived human mast cells

Jensen, Bettina M, Frandsen, Pernille M, Raaby, Ellen M, Schiøtz, Peter Oluf, Skov, Per S, Poulsen, Lars K·2014·Journal of Leukocyte Biology
DOI 10.1189/jlb.0712354
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Abstract

Different protocols exist for in vitro development of HuMCs from hematopoietic stem cells, which results in distinct mast cells regarding molecular markers and activation patterns. Here, we introduce a SR profile using immunological, neurogenic, and pharmacological stimuli to characterize cellular functionality. Mast cells were obtained from three culture protocols using two types of PBdMCs (CD34+ PBdMC or CD133+ PBdMC) and one type of CBdMC (CD133+ CBdMC). We analyzed resting cells for specific mast cell markers at protein and mRNA levels, thereby creating a molecular profile. To characterize the SR profile, we stimulated cells with anti-IgE, C3a, C5a, Substance P, or Compound 48/80 and measured the release of histamine and cytokines (IL-10, IL-13, GM-CSF, TNF-α). Molecular profiling revealed that CD133+ CBdMC expressed less chymase, FcɛRIα, and CD203c but more CD117 compared with CD34+ and CD133+ PBdMC. The SR profile for histamine release illustrated a functional heterogeneity between PBdMC and CBdMC. PBdMC released >10% histamine upon stimulation with anti-IgE, C3a, Substance P, and Compound 48/80, whereas CBdMC only reacted to C3a. Cytokine secretion was only detected after anti-IgE stimulation. Here, the SR profile identified the CD133+ PBdMC as the most active cells regarding secretion of IL-10, IL-13, GM-CSF, and TNF-α. Cells from all three culture protocols, however, produced IL-10 spontaneously at comparable levels. We recommend validating mast cell cultures by means of molecular and SR profiles to characterize the mast cells and enhance consensus among studies.

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Metadata

Authors
Jensen, B. M., Frandsen, P. M., Raaby, E. M., Schiøtz, P. O., Skov, P. S., & Poulsen, L. K.
Venue
Journal of Leukocyte Biology
Publisher
Oxford University Press (OUP)
Published
2014-06-01
Volume
95
Issue
6
Pages
893-901
ISSN
  • 0741-5400
  • 1938-3673
DOI
10.1189/jlb.0712354
Cited by
26
References
0
Language
en
Licence
academic.oup.com/pages/standard-publication-reuse-rights (vor)

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