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Phase-contrast microscopic studies using cinematographic techniques and scanning electron microscopy on IgE-mediated
M Kurosawa1, H Inamura, N Kanbe
1Department of Dermatology, Gunma University School of Medicine, Maebashi, Japan.
Insights
Cultured human mast cells, derived from umbilical cord blood, undergo exocytosis during IgE-mediated activation, releasing histamine. This finding validates their use for studying human mast cell functions.
Area of Science:
- Immunology
- Cell Biology
Background:
- Isolating primary human mast cells is challenging.
- Cultured human mast cells from umbilical cord blood offer a viable alternative.
- Investigating IgE-mediated activation mechanisms in these cells is crucial.
Purpose of the Study:
- To morphologically verify IgE-mediated activation of cultured human mast cells.
- To confirm the role of exocytosis in this process.
Main Methods:
- Human mast cells were cultured from umbilical cord blood with stem cell factor and interleukin-6.
- IgE-sensitized cells were activated with anti-IgE.
- Morphological changes were assessed using phase-contrast and scanning electron microscopy.
- Histamine release was quantified via high-performance liquid chromatography.
Main Results:
- Activated mast cells showed swelling and granule extrusion.
- Scanning electron microscopy revealed smooth bodies extruded from surface pores.
- Significant histamine release correlated with these morphological changes.
Conclusions:
- IgE-mediated histamine release from cultured human mast cells is morphologically consistent with exocytosis.
- These findings support the utility of cultured human mast cells for functional studies.
Background:
Isolating human mast cells is a laborious procedure. Recently, cultured human mast cells raised from umbilical cord blood cells have become available. It is necessary to investigate whether IgE-mediated activation of these cells is mediated by exocytosis.
Objective:
To verify IgE-mediated activation of these cultured human mast cells morphologically.
Methods:
The mast cells were raised from human umbilical cord blood cells in the presence of stem cell factor and interleukin-6. IgE-sensitized cultured human mast cells were activated by anti-IgE, and morphological changes of the cells were examined under phase-contrast microscopy using cinematographic techniques and scanning electron microscopy. Histamine release from the cells was measured with high-performance liquid chromatography.
Results:
Under the condition in which a significant histamine release was observed from the mast cells, phase-contrast microscopy showed that the cultured human mast cells became swollen and extruded granules. Scanning electron microscopy disclosed the extrusion of smooth and round bodies from pores formed on the activated mast cell surface.
Conclusion:
IgE-mediated histamine release from cultured human mast cells is accompanied by exocytosis morphologically, indicating that cultured human mast cells will help in studying the functional properties of human mast cells.