Isolation of Mature (Peritoneum-Derived) Mast Cells and Immature (Bone Marrow-Derived) Mast Cell Precursors From Mice

PLoS One. 2016 Jun 23;11(6):e0158104. doi: 10.1371/journal.pone.0158104. eCollection 2016.

Abstract

Mast cells (MCs) are a versatile cell type playing key roles in tissue morphogenesis and host defence against bacteria and parasites. Furthermore, they can enhance immunological danger signals and are implicated in inflammatory disorders like fibrosis. This granulated cell type originates from the myeloid lineage and has similarities to basophilic granulocytes, both containing large quantities of histamine and heparin. Immature murine mast cells mature in their destination tissue and adopt either the connective tissue (CTMC) or mucosal (MMC) type. Some effector functions are executed by activation/degranulation of MCs which lead to secretion of a typical set of MC proteases (MCPT) and of the preformed or newly synthesized mediators from its granules into the local microenvironment. Due to the potential accumulation of mutations in key signalling pathway components of corresponding MC cell-lines, primary cultured MCs are an attractive mean to study general features of MC biology and aspects of MC functions relevant to human disease. Here, we describe a simple protocol for the simultaneous isolation of mature CTMC-like murine MCs from the peritoneum (PMCs) and immature MC precursors from the bone marrow (BM). The latter are differentiated in vitro to yield BM-derived MCs (BMMC). These cells display the typical morphological and phenotypic features of MCs, express the typical MC surface markers, and can be propagated and kept in culture for several weeks. The provided protocol allows simple amplification of large quantities of homogenous, non-transformed MCs from the peritoneum and bone marrow-derived mast cells for cell- and tissue-based biomedical research.

MeSH terms

  • Animals
  • Biomarkers
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / metabolism*
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Separation
  • Cells, Cultured
  • Female
  • Immunophenotyping
  • Male
  • Mast Cells / cytology*
  • Mast Cells / metabolism*
  • Mice
  • Peritoneum / cytology*
  • Phenotype
  • Stem Cells / cytology*
  • Stem Cells / metabolism*

Substances

  • Biomarkers

Grant support

This study was supported by the Deutsche Forschungsgemeinschaft, www.dfg.de, grants SFB/TRR57 (P13, Q3) and HU794/10-1, and IZKF Aachen, www.izkf-aachen.de/#, grant E7-6. The sponsors had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.