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Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
Published on: July 4, 2018
Histamine release and surface CD200R1 staining as sensitive methods for assessing murine mast cell activation
1Department of Microbiology and Immunology, Uniformed Services University, 4301 Jones Bridge Road, Bethesda, MD, USA. dlarson@usuhs.mil
Journal of Immunological Methods
|March 8, 2012
Summary
Histamine release and CD200R1 expression are highly sensitive methods for measuring mast cell activation. These techniques require fewer cells than traditional assays, making them ideal for disease research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Mast cells are key players in allergic reactions and disease pathology.
- Current methods like beta-hexosaminidase activity assays need many cells, limiting their use in disease models.
Purpose of the Study:
- To compare the sensitivity of histamine release and beta-hexosaminidase activity for measuring mast cell activation.
- To introduce a novel, sensitive flow cytometry method for detecting mast cell activation.
Main Methods:
- Compared histamine release and beta-hexosaminidase activity in response to anti-IgE and ionomycin stimulation.
- Developed and validated a flow cytometry assay measuring surface CD200R1 expression on mast cells after stimulation.
Main Results:
- Histamine release detected mast cell activation with as few as 480 cells/ml, significantly more sensitive than beta-hexosaminidase (6x10^4 cells/ml).
- Flow cytometry showed increased CD200R1 expression on IgE+c-kit+ cells within 30 minutes of stimulation, using 7.5x10^5 peritoneal cells.
- Both methods work with unpurified cells and require low cell numbers.
Conclusions:
- Histamine release and CD200R1 expression via flow cytometry are highly sensitive and practical methods for assessing murine mast cell activation.
- These improved methods are well-suited for studying mast cell activation in various mouse disease models.
