Protocol for isolating immune cells from human gastric muscularis propria for single-cell analysis

STAR Protoc. 2024 Sep 20;5(3):103258. doi: 10.1016/j.xpro.2024.103258. Epub 2024 Aug 11.

Abstract

Understanding the diversity of gastrointestinal (GI) immune cells, especially in the muscularis propria, is crucial for understanding their role in the maintenance of enteric neurons and smooth muscle and their contribution to GI motility. Here, we present a detailed protocol for isolating single immune cells from the human gastric muscularis propria. We describe steps for tissue preservation, dissection, and dissociation of the muscularis propria. We then detail procedures for magnetic sorting of CD45+ cells and single-cell RNA sequencing (scRNA-seq) analysis. For complete details on the use and execution of this protocol, please refer to Chikkamenahalli et al.1.

Keywords: Cell isolation; Genomics; Immunology; Molecular Biology; RNA-seq; Sequencing; Single Cell; cell Biology.

MeSH terms

  • Cell Separation / methods
  • Gastric Mucosa* / cytology
  • Humans
  • Single-Cell Analysis* / methods
  • Stomach / cytology