Rapid isolation of mouse ILCs from murine intestinal tissues

Methods Enzymol. 2020;631:305-327. doi: 10.1016/bs.mie.2019.10.001. Epub 2019 Nov 12.


Tissue-resident immune cells like innate lymphoid cells (ILCs) are regulators of local immunity and tissue homeostasis. Similar to Natural Killer (NK) cells, ILCs express germline-encoded natural cytotoxicity receptors (NCRs) that facilitate the rapid execution of effector functions. Recent advances using transgenic animal models have further uncovered the developmental, transcriptional, epigenetic, and functional differences between members of the ILC family. Isolation of ILCs, which are particularly enriched in non-lymphoid tissues, can often be challenging and time consuming. Here, we provide a simple and rapid protocol for the isolation of NK cells and ILCs from murine intestinal tissues. This protocol is suitable for Fluorescence Activated Cell Sorting (FACS) and intracellular analysis of cytokine and transcription factor expression using flow and mass cytometry.

Keywords: ILC1; ILC2; Innate lymphoid cells; Intestine; Lamina propria; Mucosal immunology; NK cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Separation / methods*
  • Cytokines / analysis
  • Cytokines / metabolism
  • Flow Cytometry / methods
  • Gene Expression Regulation
  • Immunity, Innate
  • Intestines / immunology*
  • Lymphocytes / immunology
  • Lymphocytes / metabolism*
  • Mice
  • Transcription Factors / analysis
  • Transcription Factors / genetics


  • Cytokines
  • Transcription Factors

Grant support