Isolation of primary immune cells from fibrotic skin, esophageal, and gut tissue

J Immunol Methods. 2021 Oct:497:113107. doi: 10.1016/j.jim.2021.113107. Epub 2021 Aug 2.

Abstract

Understanding the interplay between immune and structural cells is important for studying fibrosis and inflammation; however, primary immune cell isolation from organs that are typically enriched in stromal cells, like the lung, esophagus, or gut, proves to be an ongoing challenge. In fibrotic conditions, this challenge becomes even greater as infiltrating cells become trapped in the robust extracellular matrix (ECM). This protocol details a method to isolate cells at high yield from stroma-rich organs that can be used for further analyses via flow cytometry, stimulation, or culturing. Validation of this method is confirmed by flow cytometry data assessing immune cell populations of interest. This protocol can be completed in approximately 5-6 h.

Keywords: Fibrosis; Immune cell isolation; Mucosa.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Separation*
  • Cell Survival
  • Cells, Cultured
  • Collagenases / metabolism
  • Endopeptidases / metabolism
  • Esophageal Mucosa / cytology*
  • Esophageal Mucosa / immunology
  • Esophageal Mucosa / metabolism
  • Fibrosis
  • Flow Cytometry*
  • Intestinal Mucosa / cytology*
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Skin / immunology
  • Skin / metabolism
  • Skin / pathology*
  • Time Factors
  • Trypsin / metabolism
  • Workflow

Substances

  • Biomarkers
  • Endopeptidases
  • Trypsin
  • Collagenases
  • dispase