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Updated: Dec 21, 2025

An Ex vivo Mast Cell Degranulation Assay using Crude Peritoneal Exudate Cells and Natural Antigen Stimulation
Published on: April 27, 2021
Novel reactivation and degranulation of mast cells
Huanjin Liao1, Xia Peng1, Yiqin Ge1
1Department of Laboratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
Abstract:
Mast cells (MCs) degranulation is a key process during the allergic inflammatory response. MCs release their preformed and new synthesized granules after activation. We found that granules were released partially and selectively after the activation of bone marrow-derived mouse mast cells (BMMCs). Next, we investigated the response of degranulated MCs to a new challenge. BMMCs were activated by antibody/antigen (IgE/Ag) or compound 48/80 (C48/80). The degranulated BMMCs were then reactivated by either IgE/Ag or C48/80 without time intervals. Flow cytometry was used to detect the expression of CD117, FcεRI, and intracellular granules of BMMCs, and BMMCs degranulation was detected using the β-hexosaminidase release assay. The morphology of BMMCs was observed by staining with toluidine blue. Degranulated BMMCs activated by IgE/Ag failed to respond to the same IgE/Ag challenge and released β-hexosaminidase independent of unoccupied FcεRI, but responded to C48/80. Degranulated BMMCs activated by C48/80 responded to either IgE/Ag or C48/80. These results indicated that degranulated BMMCs could be reactivated and released granule mediators again, this revealed the unique mediator releasing mechanism of degranulated MCs and their potential function in maintaining inflammation or causing hypersensitivity.
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