Developmental and Functional Assays to Study Murine and Human γδ T Cells

Methods Mol Biol. 2017:1514:257-267. doi: 10.1007/978-1-4939-6548-9_18.


The key roles played by gamma-delta (γδ) T cells in immunity to infection and tumors critically depend on their differentiation into effectors capable of secreting cytokines (such as interferon-γ or interleukin-17), and killing infected or transformed cells. Here we detail the main methods used to investigate the differentiation of γδ T cells from murine or human origin. We describe developmental assays, such as thymic organ cultures (TOCs) and coculture of progenitors cells with OP9-DL1 stomal cells, as well as functional assays typically employed to evaluate γδ T cell cytotoxicity and cytokine production.

Keywords: Cytokine production; Cytotoxicity; Effector differentiation; Intracellular FACS staining; Thymic development; Thymic organ culture (TOC); γδ T cells.

MeSH terms

  • Animals
  • Cell Differentiation / genetics*
  • Cell Differentiation / immunology
  • Humans
  • Interferon-gamma / metabolism
  • Interleukin-17 / metabolism
  • Intraepithelial Lymphocytes / cytology*
  • Intraepithelial Lymphocytes / immunology
  • Mice
  • Organ Culture Techniques / methods*
  • T-Lymphocyte Subsets / cytology*
  • T-Lymphocyte Subsets / immunology
  • Thymus Gland / cytology
  • Thymus Gland / immunology


  • Interleukin-17
  • Interferon-gamma