Mechanisms of allergy/immunology
A novel approach for studying mast cell–driven disorders: Mast cells derived from induced pluripotent stem cells

https://doi.org/10.1016/j.jaci.2021.07.027Get rights and content

Background

Mast cells (MCs) are considered the main effectors in allergic reactions and well known for their contribution to the pathogenesis of various inflammatory diseases, urticaria, and mastocytosis. To study their functions in vitro, human primary MCs are isolated directly from several tissues or differentiated from hematopoietic progenitors. However, these techniques bear several disadvantages and challenges including low proliferation capacity, donor-dependent heterogeneity, and the lack of a continuous cell source.

Objective

To address this, we developed a novel strategy for the rapid and efficient differentiation of MCs from human-induced pluripotent stem cells (hiPSCs).

Methods

A 4-step protocol for the generation of hiPSC-derived MCs, based on the use of 3 hiPSC lines, was established and validated by comparison with human skin MCs and peripheral hematopoietic stem cell–derived MCs.

Results

hiPSC-MCs share phenotypic and functional characteristics of human skin MCs and peripheral hematopoietic stem cell–derived MCs. They display stable expression of the MC-associated receptors CD117, FcεRIα, and Mas-related G protein–coupled receptor X2 and degranulate in response to IgE/anti-IgE and substance P.

Conclusions

This novel hiPSC-based approach provides a sustainable and homogeneous source for a rapid and highly productive generation of phenotypically mature, functional MCs, and its principle allows for the investigation of disease- and patient-specific MC populations.

Section snippets

Differentiation and culture of hiPSC-derived MCs

Three healthy human hiPSC lines (BIHi005-A, BIHi001-A, and BIHi001-B) were obtained from the cell bank of the Stem Cell Core facility, Berlin Institute of Health (see this article’s Online Repository and this article’s Methods section in the Online Repository at www.jacionline.org for additional information on hiPSC lines).

For differentiation of hiPSCs into functional MCs, we used a 4-step protocol (see this article’s Online Repository and this article’s Methods section in the Online Repository

hiPSCs differentiate into CD34+/CD45+ HSCs

Following our 4-step protocol for the generation of hiPSC-MCs from 3 different hiPSC lines (Fig 1), we first tested the ability of hiPSCs to differentiate into CD34+ HSCs. HSCs could be successfully differentiated from all of the 3 hiPSC lines when starting from 40 ± 7 hiPSC colonies with an average diameter of about 150 μm (Fig 2, Ai). The first appearance of round nonadherent cells was observed at day 5 (Fig 2, Aii), whereas at around day 8 in HSC differentiation, medium, large clusters of

Discussion

Here, we present a novel strategy for the differentiation of MCs from hiPSCs. Compared with previous protocols, our approach provides for a rapid and highly productive generation of phenotypically mature and functional MCs. To our knowledge, this is the first report of MCs derived from hiPSCs that are similar, in maturity and function, to hsMCs and PSCMCs.

We report a 4-step protocol for the generation of MCs from hiPSC lines, based on the use of 3 different hiPSC lines and validated by

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    This work was funded by GA2LEN (Global Allergy and Asthma European Network).

    Data sharing statement: For original data, please contact [email protected] as well as corresponding authors.

    Disclosure of potential conflict of interest: M. Maurer is or recently was a speaker and/or advisor for and/or has received research funding from Allakos, Amgen, Aralez, ArgenX, AstraZeneca, Blueprint, Celldex, Centogene, CSL Behring, FAES, Genentech, GIInnovation, Innate Pharma, Kyowa Kirin, Leo Pharma, Lilly, Menarini, Moxie, Novartis, Roche, Sanofi/Regeneron, Third HarmonicBio, UCB, and Uriach. F. Siebenhaar is or recently was a speaker and/or advisor for and/or has received research funding from Allakos, Blueprint, Celldex, Cogent Bio, Genentech, Glenmark, Moxie, Novartis, Pediapharm, Sanofi, and Uriach. The rest of the authors declare that they have no relevant conflicts of interest.

    ∗

    These authors contributed equally to this study.

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