Generation of Mast Cells from Murine Stem Cell Progenitors

Methods Mol Biol. 2020:2163:85-89. doi: 10.1007/978-1-0716-0696-4_7.

Abstract

Mouse bone marrow-derived mast cells (mBMMCs) are an invaluable tool for the study of mast cell function as they represent a primary source of mature mast cells. They can be sourced from wild-type, knockout, and transgenic mice and are used to repopulate mast cell-deficient mice. This method describes the isolation of mast cell hematopoietic progenitors from the bone marrow of mouse femurs and their subsequent culture in an IL-3-rich culture medium. After 4 weeks in culture, mBMMCs are obtained in high number and are of high purity. Assessment of their granularity by toluidine staining and IgE receptor expression by flow cytometry is also described. These cells are a useful tool in the determination of in vitro and in vivo mast cell function in innate and adaptive immunity.

Keywords: BMMC; Degranulation.; FcεRI; IgE; Mast cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Culture Media / chemistry
  • Culture Media / pharmacology
  • Interleukin-3 / analysis
  • Interleukin-3 / pharmacology
  • Mast Cells / cytology*
  • Mast Cells / metabolism
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Primary Cell Culture / methods*
  • Receptors, IgE / genetics
  • Receptors, IgE / metabolism

Substances

  • Culture Media
  • Interleukin-3
  • Receptors, IgE