A Novel Cell Culture System to Improve MRGPRX2 Research in Human Skin Mast Cells

IF 4.2 2区医学 Q2 ALLERGY

Clinical and Translational Allergy Pub Date : 2025-10-16 DOI:10.1002/clt2.70112

Zhuoran Li, Jean Schneikert, Anja Wegner, Torsten Zuberbier, Magda Babina

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引用次数: 0

Abstract

Background

MRGPRX2, a receptor central to mast cell (MC) activation and related skin diseases, is selectively expressed in skin MCs but downregulated during culture by stem cell factor (SCF) and interleukin-4 (IL-4).

Objective

To identify culture conditions that preserve MC viability while restoring MRGPRX2 expression and function.

Methods

Human skin MCs were cultured in standard or serum-free Accell medium (both with SCF). After 3 days, MRGPRX2 expression was assessed by flow cytometry, degranulation by mediator release assays, and signaling by Western blotting.

Results

Accell medium increased MRGPRX2 expression to ~1.7-fold and enhanced degranulation to Substance P and codeine. It also promoted stronger and more sustained ERK and AKT phosphorylation, while FcεRI-mediated responses were largely unaffected.

Conclusion

Short-term incubation in serum-free Accell medium restores MRGPRX2 expression and signaling in cultured skin MCs without impairing viability. This simple adjustment yields a practical and reliable model for MRGPRX2-focused studies.

Abstract Image

查看原文本刊更多论文

一种新的细胞培养系统提高人皮肤肥大细胞MRGPRX2的研究。

背景：MRGPRX2是肥大细胞（MC）活化和相关皮肤病的中枢受体，在皮肤MC中选择性表达，但在培养过程中被干细胞因子（SCF）和白细胞介素-4 （IL-4）下调。目的：寻找既能保持MC活力又能恢复MRGPRX2表达和功能的培养条件。方法：在标准或无血清Accell培养基（均含SCF）中培养人皮肤MCs。3天后，通过流式细胞术、介质释放法和Western blotting检测MRGPRX2的表达。结果：加速细胞培养基使MRGPRX2表达量增加约1.7倍，增强了对P物质和可待因的脱颗粒作用。它还促进了更强、更持久的ERK和AKT磷酸化，而fcε ri介导的反应在很大程度上不受影响。结论：在无血清Accell培养基中短期孵育可恢复培养的皮肤MCs中MRGPRX2的表达和信号传导，而不影响其生存能力。这种简单的调整为以mrgprx2为重点的研究提供了实用可靠的模型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical and Translational Allergy Immunology and Microbiology-Immunology

CiteScore

7.50

自引率

4.50%

发文量

117

审稿时长

12 weeks

期刊介绍： Clinical and Translational Allergy, one of several journals in the portfolio of the European Academy of Allergy and Clinical Immunology, provides a platform for the dissemination of allergy research and reviews, as well as EAACI position papers, task force reports and guidelines, amongst an international scientific audience. Clinical and Translational Allergy accepts clinical and translational research in the following areas and other related topics: asthma, rhinitis, rhinosinusitis, drug hypersensitivity, allergic conjunctivitis, allergic skin diseases, atopic eczema, urticaria, angioedema, venom hypersensitivity, anaphylaxis, food allergy, immunotherapy, immune modulators and biologics, animal models of allergic disease, immune mechanisms, or any other topic related to allergic disease.