Mouse bone marrow-derived mucosal mast cell model characterization

Mast cells (MCs) fall into two subpopulations with well-defined roles and characteristics: connective tissue mast cells (CTMCs) and mucosal mast cells (MMCs). While in vitro models of CTMCs exist in humans and mice, a reliable and relevant model of MMCs is still lacking. Contrary to the pioneer studies' one-step protocol, our two-step differentiation protocol replicates the two major stages of mast cell development to generate mouse bone marrow-derived mucosal mast cells (BM-MMCs) in vitro. Bone marrow cells were cultured in a complete Opti-MEM medium supplemented with IL-3 and SCF for 4 weeks to induce MC commitment before adding for one additional week both IL-9 and TFG-ß1 to promote MMC proliferation and maturation BM-MMCs were phenotypically and functionally compared to the previously established CTMC model (PCMCs). RNAseq analysis showed that BM-MMC exibit MMC typical characteristics and recapitulate gene expression observed in isolated MMCs. BM-MMCs, which have transcriptomic characteristics similar to MMCs, can be an effective tool for studying a significant number of viable MMCs during a 15-day period in vitro Overall design: RNAseq profiling of BM-MMCs (n=3) as compared with PCMCs (n=3)