Lin− CD34hi CD117int/hi FcεRI+ cells in human blood constitute a rare population of mast cell progenitors

Dahlin, Joakim S.; Malinovschi, Andreï; Öhrvik, Helena; Sandelin, Martin; Janson, Christer; Alving, Kjell; Hallgren, Jenny

doi:10.1182/blood-2015-06-650648

Cited by 105 publications

(110 citation statements)

References 42 publications

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“…Re-analysis of the respective progeny cells by flow cytometry revealed that all cells had an intermediate CD117 expression after culture and that a proportion of them had lost FcεRI surface expression ( Figure 1F and J). The loss of FcεRI expression upon in vitro culture of the human peripheral blood MCp during the same culture conditions was reported previously, 3…”

Section: Demonstration Of Human Mast Cell Progenitors In the Bone Marrowsupporting

confidence: 80%

Demonstration of human mast cell progenitors in the bone marrow

Salomonsson

Ungerstedt

Alvarado-Vázquez

et al. 2019

Allergy

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Section: Demonstration Of Human Mast Cell Progenitors In the Bone Marrowsupporting

confidence: 80%

Demonstration of human mast cell progenitors in the bone marrow

Salomonsson

Ungerstedt

Alvarado-Vázquez

et al. 2019

Allergy

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“…The total numbers of cells after enrichment were also similar regardless of the origin of the PBMCs (2.43 ± 0.10 ×10 6 from lymphocytapheresis and 2.81 ± 0.72 ×10 6 from whole blood). Mast cell progenitors (MCP) are reported to co-express CD34, CD117 and FcεRI[14]. Among CD34 + enriched cells, no differences were observed in the number of MCP obtained from either lymphocytapheresis (4.14%; N=4) or whole blood (3.48%; N=3) (Fig 1C; gating strategy shown in Supplementary Fig 1A).…”

Section: Resultsmentioning

confidence: 99%

“…Human peripheral blood has been used as a source for culturing huMCs[4, 14]. Current methods of culture tend to be time-consuming and costly.…”

Section: Discussionmentioning

confidence: 99%

An optimized protocol for the generation and functional analysis of human mast cells from CD34 + enriched cell populations

Yin

Bai

Olivera

et al. 2017

Journal of Immunological Methods

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The culture of mast cells from human tissues such a cord blood, peripheral blood or bone marrow aspirates has advanced our understanding of human mast cells (huMC) degranulation, mediator production and response to pharmacologic agents. However, existing methods for huMC culture tend to be laborious and expensive. Combining technical approaches from several of these protocols, we designed a simplified and more cost effective approach to the culture of mast cells from human cell populations including peripheral blood and cryopreserved cells from lymphocytapheresis. On average, we reduced by 30–50 fold the amount of culture media compared to our previously reported method, while the total MC number generated by this method (2.46 ± 0.63 × 106 vs. 2.4 ± 0.28 × 106, respectively, from 1.0 × 108 lymphocytapheresis or peripheral blood mononuclear blood cells [PBMCs]) was similar to previous method (2.36 ± 0.70 × 106), resulting in significant budgetary savings. In addition, we compared the yield of huMCs with or without IL-3 added to early cultures in the presence of stem cell factor (SCF) and interlukin-6 (IL-6) and found that the total MC number generated, while higher with IL-3 in the culture, did not reach statistical significance, suggesting that IL-3, often recommended in the culture of huMCs, is not absolutely required. We then performed a functional analysis by flow cytometry using standard methods and which maximized the data we could obtain from cultured cells. We believe these approaches will allow more laboratories to culture and examine huMC behavior going forward.

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“…Mast cell progenitors circulate in the blood as Lin - CD34 hi CD117 int/hi FcεRI + cells. 33 These cells express CD123 and lack CD45RA, 23,33 overlaying the CMP FcεRI+ gate in bone marrow (Supplementary Fig. 3A) that exhibit a distinct expression profile compared with mature mast cells (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We have previously reported that CD34 + CD117 int/hi FcεRI + cells in peripheral blood are committed or nearly committed MCPs. 33 Considering the CD34 + CD117 + FcεRI + phenotype of CMPs FcεRI+ cells, these cells might be expected to present a similar mast cell-forming capacity as to blood MCPs. In the current study, we showed that CMPs FcεRI+ gave rise to a higher frequency of mast cells than CMPs FcεRI- and GMPs.…”

Section: Discussionmentioning

confidence: 99%

Single-cell analysis reveals the KIT D816V mutation in hematopoietic stem and progenitor cells in systemic mastocytosis: Supplementary data

Grootens

Ungerstedt

Ekoff

et al. 2018

Preprint

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32Background: Systemic mastocytosis (SM) is a hematological disease characterized 33 by organ infiltration by neoplastic mast cells. Almost all SM patients have a mutation 34 in the gene encoding the tyrosine kinase receptor KIT causing a D816V substitution 35 and autoactivation of the receptor. Mast cells and CD34 + hematopoietic progenitors 36 can carry the mutation, however, in which progenitor cell subset the mutation arises is 37 unknown. We aimed to investigate the distribution of the D816V mutation in single 38 mast cells and single hematopoietic stem and progenitor cells. 39Methods: Fluorescence-activated single-cell index sorting and D816V mutation 40 assessment were applied to analyze mast cells and more than 10,000 CD34 + bone 41 marrow progenitors across 10 hematopoietic progenitor subsets. In vitro assays 42 verified cell-forming potential. 43Findings: We found that in SM 60-99% of the mast cells harbored the D816V 44 mutation. Despite increased frequencies of mast cells in SM patients compared with 45 control subjects, the hematopoietic progenitor subset frequencies were comparable. 46Nevertheless, the mutation could be detected throughout the hematopoietic landscape 47 of SM patients, from hematopoietic stem cells to more lineage-primed progenitors. In 48 addition, we demonstrate that FcεRI + bone marrow progenitors exhibit mast cell-49 forming potential, and we describe aberrant CD45RA expression on SM mast cells for 50 the first time. 51Interpretation: The KIT D816V mutation arises in early hematopoietic stem and 52 progenitor cells and the mutation frequency is approaching 100% in mature mast 53 cells, which express the aberrant marker CD45RA. 54 55

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Lin− CD34hi CD117int/hi FcεRI+ cells in human blood constitute a rare population of mast cell progenitors

Cited by 105 publications

References 42 publications

Demonstration of human mast cell progenitors in the bone marrow

Demonstration of human mast cell progenitors in the bone marrow

An optimized protocol for the generation and functional analysis of human mast cells from CD34 + enriched cell populations

Single-cell analysis reveals the KIT D816V mutation in hematopoietic stem and progenitor cells in systemic mastocytosis: Supplementary data

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