A Technique to Isolate Primary Mast Cells from a Murine Peritoneal Cavity
Published: August 31, 2023
Abstract
Source: Tsvilovskyy, V., et al. Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays. J. Vis. Exp. (2018).
This video demonstrates a peritoneal lavage method for isolating mature connective tissue-type mast cells from the peritoneal cavity of adult mice. Upon isolating a mixture of primary immune cells from the peritoneal cavity, selective proliferation of mast cells is performed under appropriate culture conditions.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. PMC isolation by intraperitoneal lavage Prior to the cell isolation, prepare the materials listed in Table 1. Use 8 to 14-week-old male mice for PMC isolation. Euthanize a mouse by CO2 inhalation, and confirm the death by loss of reflexes. Spray the mouse with 70% ethanol and fix it…
Disclosures
The authors have nothing to disclose.
Materials
| RPMI Medium | Thermo Scientific | 21875034 | |
| DPBS | Sigma-Aldrich | 14190144 | |
| FCS | Thermo Scientific | R92157 | |
| IL-3 | R&D Systems | 403-ML | |
| SCF | Thermo Scientific | PMC2115 | |
| Penstrep | Thermo Scientific | 15140122 | |
| Culture Flasks (25 cm) | Greiner Bio-One | 69160 | |
| 15 mL Plastic Centrifuge Tubes | Sarstedt | 6,25,54,502 | |
| Bench Centrifuge | Heraeus | Megafuge 1.0R |
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Cite This Article
A Technique to Isolate Primary Mast Cells from a Murine Peritoneal Cavity. J. Vis. Exp. (Pending Publication), e21572, doi: (2023).