Related Experiment Video
Updated: Aug 8, 2026
Analyzing the Functions of Mast Cells In Vivo Using 'Mast Cell Knock-in' Mice
Published on: May 27, 2015
Mast-cell heterogeneity: functional comparison of purified mouse cutaneous and peritoneal mast cells
D He1, S Esquenazi-Behar, N A Soter
1Dermatology Service, New York Veterans Administration Medical Center, New York 10010.
Insights
This study reveals significant functional differences between mouse cutaneous mast cells (CMC) and peritoneal mast cells (PMC). These mast cell populations exhibit distinct responses to various secretagogues, highlighting their specialized roles.
Area of Science:
- Immunology
- Cell Biology
- Allergy Research
Background:
- Mast cells are crucial immune cells involved in allergic reactions and inflammation.
- Understanding mast cell heterogeneity is key to developing targeted therapies.
- Previous research suggests potential differences between mast cell populations.
Purpose of the Study:
- To investigate the functional heterogeneity of mouse cutaneous mast cells (CMC) and peritoneal mast cells (PMC).
- To compare the responses of CMC and PMC to various secretagogues, including A23187, compound 48/80, and substance P.
- To elucidate the distinct characteristics and potential roles of different mast cell subtypes.
Main Methods:
- Isolation and purification of CMC from mouse ears and PMC from peritoneal lavage.
- Quantification of histamine content and assessment of cell purity and viability.
- Measurement of histamine release from both mast cell types in response to secretagogues and varying conditions (concentration, time, temperature, calcium dependency).
Main Results:
- Peritoneal mast cells (PMC) showed significantly higher histamine content than cutaneous mast cells (CMC).
- CMC and PMC displayed differential dose- and time-dependent histamine release patterns in response to A23187, compound 48/80, and substance P.
- Calcium dependency of histamine release varied between CMC and PMC, particularly in response to substance P.
Conclusions:
- Functional heterogeneity exists between mouse cutaneous and peritoneal mast cells.
- These distinct functional properties suggest specialized roles for CMC and PMC in different physiological and pathological contexts.
- The findings provide a basis for further research into mast cell-specific therapies.
Abstract:
To investigate the functional heterogeneity of mouse mast cells, we extracted and purified cutaneous and peritoneal mast cells from 10- to 18-week-old BALB/c mice and compared their responses to secretagogues. Cutaneous mast cells (CMC) were extracted from mouse ears after digestion with hyaluronidase and collagenase in MEM containing 25% fetal calf serum and purified on a discontinuous Percoll gradient. The histamine content of cells obtained from the 30/40% interface was 1.0 +/- 0.1 pg/cell (mean +/- SE), with a mast-cell purity of 68.6 +/- 4.4% and a viability of greater than 93%. Peritoneal mast cells (PMC) were obtained by lavage with modified Tyrode's buffer followed by purification on 22.5% and 3-9% metrizamide gradients. The histamine content of cells was 12.2 +/- 0.8 pg/cell, with a mast-cell purity of 95.9 +/- 0.6% and a viability of greater than 95%. Histamine release induced by A23187 from CMC peaked at 3.0 microM A23187 (19.1 +/- 4.2%), at 3.0 min (22.3 +/- 2.3%), and at 30 degrees C (17.6 +/- 2.6%). In contrast, histamine release from PMC peaked at 8.0 microM of A23187 (49.4 +/- 12.1%) and at 15.0 min (48.5 +/- 12.2%). Release of histamine from PMC was observed at all the temperatures tested from 22 to 45 degrees C. Histamine release from CMC and PMC induced by A23187 was calcium dependent. Histamine release induced by compound 48/80 from CMC peaked at 0.5 micrograms/ml of compound 48/80 (23.0 +/- 7.4%) and at 5.0 min incubation (16.3 +/- 2.0%), whereas release from PMC peaked at 10.0 micrograms/ml (31.9 +/- 2.6%); release from PMC was similar at all the time points examined (1-15 min). Histamine release induced by substance P (SP) from both CMC and PMC peaked at 5.0 microM (18.8 +/- 6.6% and 12.6 +/- 3.7%, respectively); however, the maximal release from CMC occurred at 3.0 min (18.2 +/- 3.2%) and from PMC at 30.0 min (11.4 +/- 2.0%). SP-induced histamine release from CMC was calcium dependent, whereas release from PMC was only partially inhibited by EDTA. This study demonstrated that functional heterogeneity exists between these two populations of mast cells.
