Detection and Isolation of Airway Mast Cell Subsets in Mouse and Human

Mast cells are heterogeneous, tissue-resident immune effector cells widely recognized to play pathobiologic roles in the development of mucosal type 2 inflammation. While mast cell progenitors can be found in peripheral blood, phenotypically mature mast cells can only be found within peripheral tissues. Here, we describe optimized tissue digestion protocols for obtaining mast cells from the murine lung and from human sinus tissue. Following tissue digestion, mast cells can be identified and sorted by flow cytometry using antibodies against defined surface markers. We also provide protocol for intracellular protease immunostaining, allowing for further characterization of mast cells.